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1
Tassios, Panayotis. "Control of transcription in embryonal carcinoma cells." Thesis, University College London (University of London), 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.283568.
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Paterno, Gary David. "X chromosome inactivation in mouse embryonal carcinoma cells." Thesis, University of Ottawa (Canada), 1985. http://hdl.handle.net/10393/4629.
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Bell, S. M. "The cellular immune response to murine embryonal carcinoma cells." Thesis, University of Oxford, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.354817.
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Sheardown, Steven Andrew. "The developmental genetics of mouse embryonal stem cells and embryonal carcinoma cells." Thesis, University of Edinburgh, 1989. http://hdl.handle.net/1842/24310.
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Rehbini, Ohoud Mohammedsabri M. "The role of high mobility nucleosomal binding protein (Hmgn2) in undifferentiated mouse epiblast carcinoma stem cells." Thesis, University of Glasgow, 2016. http://theses.gla.ac.uk/7190/.
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High mobility group nucleosome binding (HMGN) proteins belong to the superfamily of high mobility group (HMG) proteins. HMGN1 and HMGN2 are ubiquitously expressed in all vertebrates, and are most highly expressed in embryonic tissue. Moreover, HMGN1 and HMGN2 were found to be highly expressed in neural stem/progenitor cells in the mouse brain. Here, mouse embryonal carcinoma cells (P19 EC) were used as a model system to study the role of HMGN proteins in pluripotent stem cells. Previously, experiments using short interfering RNA (siRNA) technology to knockdown HMGN1 and HMGN2 have suggested that HMGN proteins are important for the expression of key pluripotent genes, Oct4, Sox2 and Nanog, in P19 EC cells (Mohan, 2012). The aim of this thesis was to develop a lentiviral system for the long term knockdown of Hmgn2, in order to investigate more fully the role of this protein in stem cell pluripotency and differentiation. Constitutive and inducible lentiviral shRNAmir systems were tested and optimized, and a constitutive system was chosen for further work. HMGN2 knockdown in undifferentiated P19 EC cells resulted in the down-regulation of Oct4 protein levels. ChIP assays showed that HMGN2 binding over the Oct4 gene was absent in HMGN2 knockdown cells. Furthermore, binding of HMGN1 at this locus was increased in the absence of HMGN2. Consistent with the reduction in Oct4 expression, levels of the active histone modification, H3K4me3, were also decreased at the Oct4 gene. These results support a role for HMGN2 in the regulation of Oct4 expression in P19 cells, and imply that HMGN2 may be important for maintaining stem cell pluripotency.
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Thompson, Alexandra Inés. "Investigation of the role of hepatic stellate cells in acute liver failure and hepatocarcinogenesis." Thesis, University of Edinburgh, 2017. http://hdl.handle.net/1842/28936.
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Introduction: Hepatic stellate cells (HSC) and myofibroblasts may be relevant stromal drivers of human hepatocellular carcinoma (HCC). It was hypothesised that targeted inhibition of αv integrin-mediated TGF-β activation, by HSC or hepatocytes, may result in reduced peri-tumoural and intra-tumoural extracellular matrix formation, and reduced hepatic carcinogenesis. The role of HSC in acute liver injury is less well characterised. It was anticipated that integrin signalling on HSC and hepatocytes might also be relevant in the acute setting. The emerging technique of intravital microscopy (IVM) allows detailed, real-time investigation of the cellular processes involved in hepatocyte injury, cell death and repair. It was hypothesised that this could be coupled with mouse models of HCC and acute liver injury, to perform sequential imaging under anaesthesia. Aims: (i) To determine the effect of targeted inhibition of αv integrins on HSC and hepatocytes, during hepatocarcinogenesis, in a mouse model of HCC. (ii) To investigate the effect of targeted inhibition of αv and other integrins on HSC, hepatocytes, and liver sinusoidal endothelial cells (LSEC), during acute liver injury, in the mouse model of paracetamol-induced liver injury. (iii) To develop IVM of the liver, via an abdominal imaging window, with optimisation of surgical and imaging techniques, to allow sequential imaging of the same animal. Methods: The diethylnitrosamine (DEN)-induced mouse model of hepatocarcinogenesis was used, and PDGFRβ-Cre;αvfl/fl and Alb-Cre;αvfl/fl mice were employed to deplete αv integrins on HSC and hepatocytes respectively. Tumours were harvested at 40 weeks post-DEN. Tumour size and number was evaluated in all animals. PDGFRβ-Cre;αvfl/fl and Alb-Cre;αvfl/fl mice were used in the paracetamol model, to investigate the role of αv integrins in acute liver injury. PDGFRβ-Cre;β8fl/fl and Alb-Cre;β 8fl/fl animals were also tested in this model. The role of integrins in liver sinusoidal endothelial cells (LSEC) during paracetamol-induced liver injury was evaluated using Cdh5-Cre mice. IVM of the liver was performed by surgical implantation of an abdominal imaging window, consisting of a titanium ring and coverslip, secured in place with a purse string suture. Fluorescent reporter mice were used to identify hepatic and vascular architecture, and other label-free microscope technologies were utilised to image collagen, lipid distribution, necrotic areas and blood flow within tissues. Results: In large cohorts of PDGFRβ-Cre;αvfl/fl, Alb-Cre;αvfl/fl, and control animals, there was no difference in mean tumour size or number, at 40 weeks. Targeted inhibition of α v integrins and β 8 integrin on hepatocytes, HSC or LSEC was not protective in paracetamol-induced liver injury. IVM of the liver can be performed on animals with HCC and throughout paracetamol-induced liver injury, to obtain high quality, real-time images of multiple cell lineages and the hepatic microenvironment. Conclusions: The role of TGF-β in HCC pathogenesis is complex and context-dependent. Targeted loss of αv integrin did not result in reduction in tumour burden in this non-cirrhotic model of HCC. IVM of the liver is a powerful tool to quantify inflammatory infiltrates and assessment of vascular remodelling throughout the course of acute liver injury and regeneration, providing insights into the biological processes determining recovery.
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Aghsani, Parisa. "Effects of a plant extract from Ruptiliocarpon caracolito on the growth and differentiation of P19 mouse embryonal carcinoma cells." Thesis, University of Ottawa (Canada), 2004. http://hdl.handle.net/10393/26560.
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Present data indicate that the Ruptiliocarpon caracolito extract exerted significant anti-proliferative activity against P19 EC cells, in the absence of toxicity and significant alterations in the differentiation status of the cells. The focus of this study was to determine whether the proliferation of P19 EC cells would decrease in response to Ruptiliocarpon caracolito extract and whether the anti-proliferative activity, if observed, would be accompanied by alterations in the cell differentiation status. According to the data obtained in the present study, the plant extract exerted a significant anti-proliferative activity against the P19 EC cell line. This activity was exhibited in a dose-related fashion. Amongst the doses tested (5--70mug/ml), concentrations ranging from 30--70mug/ml showed a significant anti-proliferative activity (P < 0.001). The highest growth reduction activity of the extract was noticed at a concentration of 50mug/ml of culture medium. Viability studies indicated that the decrease in cell proliferation was not secondary to the toxicity of the extract. (Abstract shortened by UMI.)
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Mejetta, Stefania 1984. "1)Jarid2 regulates mouse epidermal stem cell activation and differentiation ; 2)Tumor heterogeneity and metastasis-initiation in human squamous cell carcinoma." Doctoral thesis, Universitat Pompeu Fabra, 2013. http://hdl.handle.net/10803/283482.
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Jarid2 is required for the genomic recruitment of the polycomb
repressive complex-2 (PRC2) in embryonic stem cells. However, its
specific role during late development and adult tissues remains largely
uncharacterized. In this first part of my thesis, we show that deletion of
Jarid2 in mouse epidermis reduces the proliferation and potentiates the
differentiation of postnatal epidermal progenitors, without affecting
epidermal development. In neonatal epidermis, Jarid2 deficiency
reduces H3K27 trimethylation, a chromatin repressive mark, in
epidermal differentiation genes previously shown to be targets of the
PRC2. However, in adult epidermis Jarid2 depletion does not affect
interfollicular epidermal differentiation but results in delayed hair
follicle (HF) cycling as a consequence of decreased proliferation of HF
stem cells and their progeny. We conclude that Jarid2 is required for the
scheduled proliferation of epidermal stem and progenitor cells
necessary to maintain epidermal homeostasis.
Several human and mouse solid tumors, including squamous cell
carcinomas (SCC), contain a population of Cancer Stem Cells (CSCs).
CSCs are characterized by their unique ability to initiate and propagate
the tumor; however, very little is known about their capacity to
disseminate to distant organs and give rise to metastasis. CSCs display a
great functional and molecular heterogeneity, and it has been proposed
that different CSC subclones might exist to either maintain the primary
tumor or to metastasize in distant sites. However, the identity of these
heterogeneous populations of CSCs, as well as their molecular and
functional characteristics for most type of tumors remains to be
elucidated.
Using a novel xenograft system that we have developed to study human
head and neck squamous cell carcinoma, we have identified a labelretaining
(LRC) population inside the cancer stem cell pool defined by
the high expression of CD44 and high activity of Aldh1. Unexpectedly,
tumor LRC harbor poor initiating potential, and are more sensitive to
chemotherapy than their proliferating counterparts.
Intriguingly, tumor LRCs are defined by a unique transcriptome
signature previously linked with bone and lung identity, two major sites
of SCC metastasis, suggesting they might be involved in the
colonization of distant tissues by SCC tumors.
We have also identified surface molecules, including CD36 and CD37,
that are uniquely expressed by tumor LRCs, that can be used as
surrogate markers to isolate and characterize them from primary human
SCCs.
Based on this signature, we could demonstrate that the presence or
absence of this population in the primary tumor of a large cohort of
patients with cutaneous SCC is highly predictive of the metastatic
occurrence. In addition, several markers exclusively expressed by tumor
LRCs can be targeted with drugs currently in clinical trials for the
treatment of other diseases. We are testing whether some of these
therapeutical strategies are effective to preventing or reducing the
metastatic potential of SCC tumors.Jarid2 es necesario para la localización genómica del complejo represor polycomb repressive complex-2 (PRC2) en células stem embrionarias. Sin embargo, la función de Jarid2 en las últimas fases del desarrollo embrionario y su papel en la función de los tejidos adultos no ha sido aún caracterizada en profundidad. En esta primera parte de mi tesis doctoral, mostramos que la deleción de Jarid2 en la piel de ratón no afecta al desarrollo de la epidermis, pero reduce la proliferación y potencia la diferenciación de las células progenitoras epidermales en neonatos. La piel de los ratones neonatos Jarid2-KO muestra niveles reducidos de la marca represora de la cromatina, H3K27me3, en genes necesarios para la diferenciación de las células progenitoras. En cambio, en piel adulta la depleción de Jarid2 no afecta la diferenciación de la epidermis, pero sí que resulta en una reducción del número de células stem activas de los folículos pilosos, lo que desemboca en el retraso del crecimiento de los folículos. Por lo tanto, nuestros resultados demuestran que Jarid2 es necesario para la activación y diferenciación de diferentes células stem del compartimento queratinocítico de la piel necesarios para mantener la homeostasis epidermal. Diversos tipos de tumores sólidos humanos y de ratón, incluyendo carcinomas de células escamosas (SCCs del inglés: Squamous Cell Carcinomas), contienen una población de células madre cancerosas (CSCs del inglés Cancer Stem Cells). Las CSCs se caracterizan porque pueden iniciar y propagar el tumor; sin embargo, se conoce muy poco sobre su capacidad de alcanzar órganos lejos del tumor primario y de formar metastasis. Las CSCs pueden ser muy heterogéneas tanto a nivel funcional como molecular, y se ha propuesto que podrían existir diferentes subclones sea para mantener el tumor primario, sea para formar metástasis. No obstante, no se conoce por ahora ni la identidad de estas poblaciones heterogéneas de CSCs, ni sus características a nivel funcional o molecular. Usando un nuevo sistema de xenoinjerto que hemos desarrollado en nuestro laboratorio para estudiar SCC de cabeza y cuello, hemos identificado una población que es capaz de retener el marcaje con el tiempo (LRC de inglés: Label-retaining Cells), dentro de la población total de CSSs, definidas como células dentro del tumor que muestran alta expression de CD44 y alta actividad de Aldh1. En contra de lo que esperábamos, las LRC del tumor tienen dificultad para iniciar tumores por sí solas y son más sensibles a tratamientos de quimioterapia cuando las comparamos con otras células más proliferativas. Por otra parte, las LRC del tumor se pueden definir con un transcriptoma único que ha sido relacionado anteriormente con hueso y pulmón, que son dos de los órganos donde los SCC forman metástasis preferentemente. Esto sugiere que podrían estar involucradas en la colonización de órganos alejados del SCC primario. Hemos identificado también moléculas de superficie, incluyendo CD36 y CD37, que se expresan exclusivamente en las LRC de tumor y que se pueden usar como marcadores para aislar y caracterizar las LRC de SCCs primarios humanos. Basándonos en estos marcadores, hemos podido demostrar que la presencia o no de esta población en el tumor primario predice la formación de metástasis en pacientes con SCC cutáneos. Además, diversos marcadores que hemos identificado como únicos en LRC de tumor, son diana de fármacos ya usados en la actualidad en ensayos clínicos para tratamiento de otras enfermedades. En la actualidad estamos probando si alguno de estos tratamientos puede ser efectivo para prevenir o reducir el potencial de formar metástasis en SCC.
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Zeng, Yi [Verfasser], and Stefan [Akademischer Betreuer] Endres. "Gene expression profiles of T cells after adoptive transfer in a mouse model of pancreatic carcinoma / Yi Zeng ; Betreuer: Stefan Endres." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2017. http://d-nb.info/1126407313/34.
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Hall, Charles. "Ex vivo reprogramming of tumor-reactive immune cells from FVBN202 mice bearing lung metastatic mammary carcinoma: an immunotherapeutic opportunity revealed against recurrence." VCU Scholars Compass, 2013. http://scholarscompass.vcu.edu/etd/3176.
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Metastatic breast cancer treatment has seen few advances in recent years, yet treatment resistance continues to rise, causing disease recurrence. A pilot study was performed to determine the efficacy of ex vivo expansion and reprogramming of tumor-reactive immune cells from experimental metastatic tumor-sensitized mice. Also, phenotypic changes in tumors due to metastasis or tumor microenvironment influences were characterized. Metastatic neu+ mouse mammary carcinoma (mMMC) and its distant relapsing neu-antigen-negative variant (mANV) were investigated in FVBN202 mice. Tumor-reactive central memory CD8+ T cells and activated NK/NKT cells were successfully reprogrammed and expanded during 6-day expansion from mMMC- and/or mANV-sensitized mice, resulting in tumor-specific cytotoxicity. mMMC exhibited a flexible neu-expression pattern and acquired stem-like, tumorigenic phenotype following metastasis while mANV remained stable except decreased tumorigenicity. Myeloid-derived suppressor cell (MDSC) levels were not increased. Adoptive cellular therapy (ACT) with reprogrammed tumor-reactive immune cells may prove effective prophylaxis against metastatic or recurrent breast cancer.
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Benaduce, Ana Paula. "UV-Induced Melanoma Mouse Model Dependent on Endothelin 3 Over-Expression." FIU Digital Commons, 2014. http://digitalcommons.fiu.edu/etd/1613.
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Melanoma is one of the most aggressive types of cancer. It originates from the transformation of melanocytes present in the epidermal/dermal junction of the human skin. It is commonly accepted that melanomagenesis is influenced by the interaction of environmental factors, genetic factors, as well as tumor-host interactions. DNA photoproducts induced by UV radiation are, in normal cells, repaired by the nucleotide excision repair (NER) pathway. The prominent role of NER in cancer resistance is well exemplified by patients with Xeroderma Pigmentosum (XP). This disease results from mutations in the components of the NER pathway, such as XPA and XPC proteins. In humans, NER pathway disruption leads to the development of skin cancers, including melanoma. Similar to humans afflicted with XP, Xpa and Xpc deficient mice show high sensibility to UV light, leading to skin cancer development, except melanoma. The Endothelin 3 (Edn3) signaling pathway is essential for proliferation, survival and migration of melanocyte precursor cells. Excessive production of Edn3 leads to the accumulation of large numbers of melanocytes in the mouse skin, where they are not normally found. In humans, Edn3 signaling pathway has also been implicated in melanoma progression and its metastatic potential. The goal of this study was the development of the first UV-induced melanoma mouse model dependent on the over-expression of Edn3 in the skin. The UV-induced melanoma mouse model reported here is distinguishable from all previous published models by two features: melanocytes are not transformed a priori and melanomagenesis arises only upon neonatal UV exposure. In this model, melanomagenesis depends on the presence of Edn3 in the skin. Disruption of the NER pathway due to the lack of Xpa or Xpc proteins was not essential for melanomagenesis; however, it enhanced melanoma penetrance and decreased melanoma latency after one single neonatal erythemal UV dose. Exposure to a second dose of UV at six weeks of age did not change time of appearance or penetrance of melanomas in this mouse model. Thus, a combination of neonatal UV exposure with excessive Edn3 in the tumor microenvironment is sufficient for melanomagenesis in mice; furthermore, NER deficiency exacerbates this process.
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O'Neill, Vincent John. "Characterisation of a novel multi-tissue tumour suppressor gene in mouse." Thesis, University of Glasgow, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.366186.
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Moilanen, J. (Jyri). "Functional analysis of collagen XVII in epithelial cancers and a mouse model." Doctoral thesis, Oulun yliopisto, 2016. http://urn.fi/urn:isbn:9789526211695.
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Abstract Basement membranes (BM) underlie epithelia and endothelia and surround many tissues. In cutaneous BM epithelial cells are attached to the stroma via multiprotein complexes called hemidesmosomes (HD). Collagen XVII and integrin α6β4 are components of HD and they bind to laminin 332, a component of anchoring filaments, extracellularly. The main interest of this study is the function of collagen XVII and its interactions with these proteins. What is known about the function of collagen XVII is mostly derived from its role as an adhesive component in cutaneous HD. Here we demonstrate for the first time that collagen XVII is expressed by podocytes in the human and murine glomerulus and that mutant mice lacking collagen XVII in addition to small size, blisters and diffuse hair loss, also have deficient glomerular development and a high mortality rate. We also show for the first time at the protein level that collagen XVII is expressed, and probably has a functional interaction with laminin 332, in normal colon epithelia. We demonstrate that collagen XVII is expressed by the invasive cells of human colorectal carcinoma (CRC) samples and its immunostaining is increased in metastasis in CRC. The higher proportion of collagen XVII positive tumor cells correlates with decreased disease-free survival and cancer-specific survival times and we also suggest a functional interaction between collagen XVII and laminin 332 in CRC. Previous studies have suggested that collagen XVII participates in keratinocyte migration by affecting the correlation of HD disassembly and assembly, its expression is increased in squamous cell carcinoma (SCC) and it may have a role in cell adhesion and migration in SCC carcinogenesis. Here we demonstrate upregulated collagen XVII, integrin β4 and laminin γ2 expression in actinic keratosis, Bowen’s disease and SCC. The expression of collagen XVII was increased with a high degree of variation, especially in samples taken from areas where SCC is particularly invasive. We also demonstrate in the SCC-25 cell line that lack of collagen XVII or integrin β4 severely disrupts the adhesion, migration and invasivity of these cells. Taken together, in this study we show that collagen XVII is needed for normal glomerular development, is expressed in normal colon epithelia and participates in CRC and SCC carcinogenesis together with laminin 332 and integrin β4Tiivistelmä Tyvikalvot sijaitsevat epiteelin ja endoteelin alla ja ympäröivät monia kudoksia. Ihon tyvikalvossa epiteelisoluja alla olevaan verinahkaan kiinnittää rakenne, jota kutsutaan hemidesmosomiksi (HD). Kollageeni XVII ja integriin α6β4 ovat HD:n rakenneproteiineja. Ne kiinnittyvät solun ulkopuolella laminiin 332 nimiseen proteiiniin, joka muodostaa ankkurifilamentit. Kollageeni XVII ilmentyminen ja toiminta yhdessä näiden kahden proteiinin kanssa on tämän tutkimuksen keskeisin kohde. Valtaosa tutkimuksista, jotka käsittelevät kollageeni XVII:ää, koskevat sen toimintaa ihon keratinosyyteissä. Tässä tutkimuksessa osoitimme ensi kertaa, että hiiren ja ihmisen munuaiskerästen podosyyttisolut ilmentävät kollageeni XVII. Geenimanipuloidut hiiret, joilta kollageeni XVII oli poistettu, olivat pieniä, kehittivät rakkuloita ja karvattomuutta, niillä oli korkea kuolleisuus ja niiden munuaiskerästen kehitys oli häiriintynyt. Kollageeni XVII esiintymistä proteiinitasolla, sekä mahdollista toiminnallista yhteyttä laminiin 332:een, ei aiemmin ole osoitettu paksusuolen epiteelissä. Havaitsimme, että paksu- ja peräsuolen adenokarsinooman (CRC) invasiivinen solukko ilmentää kollageeni XVII:ää, kollageeni XVII esiintyminen on merkittävän voimakasta CRC:n metastasoinnin yhteydessä ja lisääntynyt kollageeni XVII esiintyminen lyhentää syöpävapaata aikaa ja heikentää syöpäspesifistä selviytymistä. Myös CRC:ssä kollageeni XVII toiminta voi liittyä laminiini 332:een. Aiempien tutkimusten mukaan kollageeni XVII osallistuu keratinosyyttien migratioon vaikuttamalla toimivien HD:ien määrään. Sen määrän on havaittu olevan korkeampi okasolusyövässä (SCC) ja sen on ehdotettu osallistuvan syöpäsolujen adheesioon ja migraatioon SCC:n kehittyessä. Me osoitimme kohonneen kollageeni XVII, integriini β4 ja laminiini γ2 ilmenemisen aktiinisessa keratoosissa, Bowenin taudissa sekä SCC:ssä. Kollageeni XVII määrä oli korkea, mutta vaihteli paljon, sekä hiiren että ihmisen invasiivisilla SCC alueilla. Havaitsimme myös SCC-25 solulinjalla, että kollageeni XVII tai integriini β4 puutos häiritsee vakavasti solujen adheesiota, migraatiota ja invaasiota. Yhteenvetona tässä työssä osoitimme, että kollageeni XVII:ää tarvitaan munuaiskerästen kehittymisessä, sitä esiintyy paksusuolen epiteelissä, ja että kollageeni XVII osallistuu CRC:n ja SCC:n kehittymiseen yhdessä integriini β4:n ja laminiini 332:n kanssa
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Fair, Joel Vincent. "Gli2 Accelerates Cardiac Progenitor Gene Expression During Mouse Embryonic Stem Cell Differentiation." Thesis, Université d'Ottawa / University of Ottawa, 2014. http://hdl.handle.net/10393/31579.
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The Hedgehog (HH) signalling pathway and its primary transducer, GLI2, regulate cardiomyogenesis in vivo and in differentiating P19 embryonal carcinoma (EC) cells. To further assess the role of HH signalling during mouse embryonic stem (mES) cell differentiation, we studied the effects of GLI2 overexpression during mES cell differentiation. GLI2 overexpression resulted in temporal enhancement of cardiac progenitor genes, Mef2c and Nkx2-5, along with enhancement of Tbx5, Myhc6, and Myhc7 in day 6 differentiating mES cells. Mass spectrometric analysis of proteins that immunoprecipitate with GLI2 determined that GLI2 forms a complex with BRG1 during mES cell differentiation. Furthermore, modulation of HH signalling during P19 EC cell differentiation followed by chromatin immunoprecipitation with an anti-BRG1 antibody determined that HH signalling regulates BRG1 enrichment on Mef2c. Therefore, HH signalling accelerates cardiac progenitor gene expression during mES cell differentiation potentially by recruiting a chromatin remodelling factor to at least one cardiac progenitor gene.
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Mlynarek, Marcin Aleksander. "Proteomics and the identification of serum biomarkers in a mouse model of oral squamous cell carcinoma." Thesis, McGill University, 2006. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=101731.
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Objective. To establish a clinically relevant model for the identification of protein serum biomarkers for oral squamous cell carcinoma, and to identify specific candidate proteins.Methods. Samples of oral cancer and adjacent normal tissue were obtained and were transplanted orthotopically into tongues of immunocompromised mice. When the mice lost 20% of their weight, they were sacrificed by exsanguinations. The serum was analyzed by two separate protocols: DIGE/MALDI and MudPIT/LC/ESI. Preliminary validation was conducted on an established cancer marker.
Results. We identified over one hundred proteins as being differentially expressed between control and cancer-bearing mice (p<0.05); including EGFR, cytokeratin 10, gelsolin, titin, vitronectin, retinoblastoma protein family, bullous pemphigoid antigen, and clusterin.
Conclusion. We report a proteomic approach for the identification of serum biomarkers of oral cancer using an orthotopic mouse model. We identified several proteins that can be exploited as potential markers for diagnosis of oral squamous cell carcinoma.
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Gallas, Alyssa L. "Lung tumors formed in the TGFΒRII conditional knockout mouse are the result of metastasis from the spontaneous tumor in the anorectal transition zone." University of Cincinnati / OhioLINK, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1406808954.
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Ruocco, Margherita <1981>. "Generation and characterization of mouse models of Small cell lung cancer and Basal cell carcinoma for the preclinical evaluation of new therapies." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5623/.
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Background. Human small cell lung cancer (SCLC) accounting for approximately 15-20% of all lung cancers, is an aggressive tumor with high propensity for early regional and distant metastases. Although the initial tumor rate response to chemotherapy is very high, SCLC relapses after approximately 4 months in ED and 12 months in LD.
Basal cell carcinoma (BCC) is the most prevalent cancer in the western world, and its incidence is increasing worldwide. This type of cancer rarely metastasizes and the death rate is extraordinary low. Surgery is curative for most of the patients, but for those that develop locally advanced or metastatic BCC there is currently no effective treatment.
Both types of cancer have been deeply investigated and genetic alterations, MYCN amplification (MA) among the most interesting, have been found. These could become targets of new pharmacological therapies.
Procedures. We created and characterized novel BLI xenograft orthotopic mouse models of SCLC to evaluate the tumor onset and progression and the efficacy of new pharmacological strategies. We compared an in vitro model with a transgenic mouse model of BCC, to investigate and delineate the canonical HH signalling pathway and its connections with other molecular pathways.
Results and conclusions. The orthotopic models showed latency and progression patterns similar to human disease. Chemotherapy treatments improved survival rates and validated the in vivo model. The presence of MA and overexpression were confirmed in each model and we tested the efficacy of a new MYCN inhibitor in vitro.
Preliminar data of BCC models highlighted Hedgehog pathway role and underlined the importance of both in vitro and in vivo strategies to achieve a better understanding of the pathology and to evaluate the applicability of new therapeutic compounds
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Saulnier, Ronald B. "The role of extracellular matrix and growth factors in anchorage-independent growth of a mouse mammary carcinoma cell line." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp04/nq20587.pdf.
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Calcagnì, Alessia. "Analysis of TFEB function in Ksp-Cadherin16 CRE mouse lines to model a particular type of renal cell carcinoma." Thesis, Open University, 2016. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.701362.
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TFE-fusion renal cell carcinomas (TFE-fusion RCCs) are caused by chromosomal translocations that lead to the overexpression of the TFE3 and TFEB genes (Kauffman et aI, 2014). The mechanisms causing kidney tumour development starting from TFE3/TFEB gene overexpression, remain largely uncharacterized and effective targeted therapies are yet to be identified, hence the need to model these diseases in an experimental animal system (Kauffman et al, 2014). Here we show that kidney-specific TFEB overexpression, in both constitutive and inducible conditional transgenic mouse lines, resulted in a phenotype characterized by renal clear cells, multi-layered basement membranes, severe cystic pathology, and ultimately papillary carcinomas with hepatic metastases. These features closely recapitulate the phenotype observed in both TFEB- and TFE3-mediated human kidney tumors. Analysis of kidney samples from these mice revealed both transcriptional induction of genes belonging to the WNT pathway and enhanced WNT βcatenin signalling. The use of specific WNT signalling inhibitors normalized the proliferation rate of primary kidney cells derived from transgenic mice and significantly rescued the disease phenotype in the mouse model. These data shed new light on the mechanisms underlying TFE-fusion RCCs and suggest a possible therapeutic strategy based on the inhibition of the WNT pathway.
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Hübner, Doreen, Christiane Rieger, Ralf Bergmann, Martin Ullrich, Sebastian Meister, Marieta Toma, Ralf Wiedemuth, et al. "An orthotopic xenograft model for high-risk non-muscle invasive bladder cancer in mice: influence of mouse strain, tumor cell count, dwell time and bladder pretreatment." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2018. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-231536.
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Background
Novel theranostic options for high-risk non-muscle invasive bladder cancer are urgently needed. This requires a thorough evaluation of experimental approaches in animal models best possibly reflecting human disease before entering clinical studies. Although several bladder cancer xenograft models were used in the literature, the establishment of an orthotopic bladder cancer model in mice remains challenging.
Methods
Luciferase-transduced UM-UC-3LUCK1 bladder cancer cells were instilled transurethrally via 24G permanent venous catheters into athymic NMRI and BALB/c nude mice as well as into SCID-beige mice. Besides the mouse strain, the pretreatment of the bladder wall (trypsin or poly-L-lysine), tumor cell count (0.5 × 106–5.0 × 106) and tumor cell dwell time in the murine bladder (30 min – 2 h) were varied. Tumors were morphologically and functionally visualized using bioluminescence imaging (BLI), magnetic resonance imaging (MRI), and positron emission tomography (PET).
Results
Immunodeficiency of the mouse strains was the most important factor influencing cancer cell engraftment, whereas modifying cell count and instillation time allowed fine-tuning of the BLI signal start and duration – both representing the possible treatment period for the evaluation of new therapeutics. Best orthotopic tumor growth was achieved by transurethral instillation of 1.0 × 106 UM-UC-3LUCK1 bladder cancer cells into SCID-beige mice for 2 h after bladder pretreatment with poly-L-lysine. A pilot PET experiment using 68Ga-cetuximab as transurethrally administered radiotracer revealed functional expression of epidermal growth factor receptor as representative molecular characteristic of engrafted cancer cells in the bladder.
Conclusions
With the optimized protocol in SCID-beige mice an applicable and reliable model of high-risk non-muscle invasive bladder cancer for the development of novel theranostic approaches was established.
21
Hübner, Doreen, Christiane Rieger, Ralf Bergmann, Martin Ullrich, Sebastian Meister, Marieta Toma, Ralf Wiedemuth, et al. "An orthotopic xenograft model for high-risk non-muscle invasive bladder cancer in mice: influence of mouse strain, tumor cell count, dwell time and bladder pretreatment." BioMed Central, 2017. https://tud.qucosa.de/id/qucosa%3A30688.
Full textAbstract:
Background
Novel theranostic options for high-risk non-muscle invasive bladder cancer are urgently needed. This requires a thorough evaluation of experimental approaches in animal models best possibly reflecting human disease before entering clinical studies. Although several bladder cancer xenograft models were used in the literature, the establishment of an orthotopic bladder cancer model in mice remains challenging.
Methods
Luciferase-transduced UM-UC-3LUCK1 bladder cancer cells were instilled transurethrally via 24G permanent venous catheters into athymic NMRI and BALB/c nude mice as well as into SCID-beige mice. Besides the mouse strain, the pretreatment of the bladder wall (trypsin or poly-L-lysine), tumor cell count (0.5 × 106–5.0 × 106) and tumor cell dwell time in the murine bladder (30 min – 2 h) were varied. Tumors were morphologically and functionally visualized using bioluminescence imaging (BLI), magnetic resonance imaging (MRI), and positron emission tomography (PET).
Results
Immunodeficiency of the mouse strains was the most important factor influencing cancer cell engraftment, whereas modifying cell count and instillation time allowed fine-tuning of the BLI signal start and duration – both representing the possible treatment period for the evaluation of new therapeutics. Best orthotopic tumor growth was achieved by transurethral instillation of 1.0 × 106 UM-UC-3LUCK1 bladder cancer cells into SCID-beige mice for 2 h after bladder pretreatment with poly-L-lysine. A pilot PET experiment using 68Ga-cetuximab as transurethrally administered radiotracer revealed functional expression of epidermal growth factor receptor as representative molecular characteristic of engrafted cancer cells in the bladder.
Conclusions
With the optimized protocol in SCID-beige mice an applicable and reliable model of high-risk non-muscle invasive bladder cancer for the development of novel theranostic approaches was established.
22
Pérez, lanzón María. "Modeling Hormone Receptor Positive Breast Cancer in Immunocompetent Mice Blocking tumor-educated MSC paracrine activity halts osteosarcoma progression Organoids for Modeling Genetic Diseases. In: International Review of Cell and Molecular Biology A preclinical mouse model of osteosarcoma to define the extracellular vesicle-mediated communication between tumor and mesenchymal stem cells Failure of immunosurveillance accelerates aging The metabolomic signature of extreme longevity: Naked mole rats versus mice Lurbinectedin synergizes with immune checkpoint blockade to generate anticancer immunity Laminin-binding integrins are essential for the maintenance of functional mammary secretory epithelium in lactation Immunoprophylactic and immunotherapeutic control of hormone receptor-positive breast cancer." Thesis, université Paris-Saclay, 2021. http://www.theses.fr/2021UPASL019.
Full textAbstract:
Les progrès de la recherche sur le cancer du sein dépendent de la disponibilité d’outils appropriés, comme les lignées cellulaires qui peuvent être implantées chez des souris immunocompétentes. La souche de souris C57Bl/6 est la plus étudiée et c’est la seule pour laquelle certaines variantes génétiques sont disponibles. Étant donné qu'aucune lignée cellulaire de carcinome mammaire à récepteurs hormonaux positifs de souche C57Bl/6 n'est disponible, nous avons décidé d'établir des lignées cellulaires de ce type. Nous avons induit des cancers du sein chez des souris C57BL/6 femelles en utilisant un analogue synthétique de la progestérone combiné à un agent endommageant l'ADN. Des lignées cellulaires ont été établies à partir de ces tumeurs et sélectionnées pour leur positivité au niveau du double récepteur (estrogène + progestérone), ainsi que pour leur transplantabilité chez les femelles C57BL/6. Parmi plusieurs lignées, une lignée cellulaire, que nous avons appelée MD5, remplissait ces critères et a permis l'établissement de tumeurs mal différenciées et très prolifératives. Ces tumeurs ont réduit leur croissance (sans toutefois régresser) lors du traitement par des antagonistes des récepteurs d’œstrogènes, ainsi que par une chimiothérapie à base d'anthracylines. Cependant, ce dernier effet n'a pas été influencé par la déplétion des lymphocytes T et, en outre, ces tumeurs n'ont pas répondu au blocage de PD-1, ce qui suggère que les tumeurs MD5 sont immunologiquement froides. En conclusion, les cellules MD5, dérivées des animaux C57BL/6, constituent un modèle de cancer du sein à récepteurs hormonaux positifs de mauvais pronosticProgress in breast cancer research relies on the availability of suitable cell lines that can be implanted in immunocompetent laboratory mice. The best explored mouse strain, C57Bl/6, is also the only one for which multiple genetic variants are available. Driven by the fact that no hormone receptor-positive C57Bl/6-derived mammary carcinoma cell lines are available, we decided to establish such cell lines. Breast cancers were induced in female C57BL/6 mice using a synthetic progesterone analogue combined with a DNA damaging agent. Cell lines were established from these tumors and selected for dual (estrogen + progesterone) receptor positivity, as well as transplantability into C57BL/6 females. One cell line, which we called MD5,fulfilled these criteria and allowed for the establishment of poorly differentiated, highly proliferative, immune cold tumors. Such tumors reduced their growth (though did not regress) upon treatment with estrogen receptor antagonists, as well as with anthracyline-based chemotherapy. However, the latter effect was not influenced by T cell depletion and MD tumors failed to respond to PD-1 blockade, suggesting that they are immunologically cold. In conclusion, C57BL/6-derived MD5 cells constitute a model of poor prognosis hormone receptor-positive breast cancer
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L'Hermitte, Antoine. "Rôle de LECT2 dans le microenvironnement immunitaire au cours de la cancérogènese hépatique." Thesis, Université Paris-Saclay (ComUE), 2016. http://www.theses.fr/2016SACLS360/document.
Full textAbstract:
Le carcinome hépatocellulaire (CHC) est la deuxième cause de mortalité par cancer dans le monde. Plusieurs études attestent du rôle du microenvironnement tumoral (MET) comme acteur fondamental de la carcinogenèse. A l’aide de modèles murins mimant un sous groupe de CHC fréquent, notre équipe avait identifié la molécule LECT2 comme un effecteur moléculaire important du MET dans le contrôle de l’agressivité tumorale.L'objectif de ma thèse a été d’adresser le rôle fonctionnel de LECT2 dans le microenvironnement immunitaire au cours du CHC.A l’aide de modèles murins, nous observons que l’absence de LECT2 entraine une accumulation importante de cellules myéloïdes dans le MET. Nous montrons que ces cellules myéloïdes sont immatures, arborent des capacités immunosuppressives puissantes vis-à-vis des lymphocytes T et ont un programme transcriptionnel permettant une action promotrice de tumeurs. De façon intéressante, l’accumulation de ces acteurs dans le microenvironnement est associée à l’émergence de nodules tumoraux indifférenciés exprimant des marqueurs de transition épithélio-mésenchymateuse/cellules progénitrices/métastases.D’un point de vue mécanistique, nous avons démontré une perte de différenciation plus importante des hépatocytes en absence de LECT2 dans des conditions d’activation de la signalisation β-caténine. Nous montrons également par des expériences de co-culture que les cellules myéloïdes infiltrant les tumeurs en absence de LECT2 ont une forte capacité à induire une perte de différenciation des hépatocytes.Enfin, nous avons analysé l'expression de LECT2 dans une vaste cohorte d’échantillons humains de CHC. Nous montrons que la diminution d’expression de LECT2 corrèle fortement avec 1)- la présence d’invasion vasculaire, 2)- la perte de différenciation des hépatocytes tumoraux et 3)- la présence d’infiltrats inflammatoires.L’ensemble de ces données démontre que LECT2 agit comme un régulateur essentiel dans la cancérogénèse hépatique à travers son action double sur les hépatocytes et sur la fonction des cellules myéloïdes infiltrant les tumeurs. Ainsi, ces travaux identifient LECT2 comme un biomarqueur potentiel ouvrant de nouvelles perspectives de traitement du CHCHepatocellular carcinoma (HCC) is the second cause of cancer-rel ated death worldwide. Several studies highlighted the tumor microenvironment (TEM) as a key player in cancer from initiation to progression steps of tumorigenesis. Using relevant HCC mouse models, our team identified the chemokine-like LECT2 as a critical actor of liver TEM in the control of tumor aggressiveness.The aim of my thesis was to address functionally the role of LECT2 in the immune microenvironment during HCC.Using mouse models, we observed that the absence of LECT2 induces a significant accumulation of myeloid cells in the TEM. We showed that these myeloid cells were immature, harbored strong immunosuppressive capabilities on T cells and expressed a transcriptional program sustaining tumor progression. Interestingly, the accumulation of these actors in the microenvironment is associated with the emergence of poorly differentiated tumor nodules expressing epithelial-to-mesenchymal transition / progenitor / metastasis markers.Mechanistically, we demonstrated that LECT2-deficient hepatocytes in the context of β-catenin activation were able to perform EMT like WT hepatocytes do after TGF-β1 challenge. In co-culture experiments, we demonstrated that tumor-infiltrating myeloid cells in the absence of LECT2 have a strong ability to induce hepatocyte EMT.Finally, we analyzed the expression of LECT2 in a vast cohort of HCC liver samples and found that downregulation of LECT2 expression strongly correlates with 1) - the presence of vascular invasion, 2) – histological grade and 3) - the presence of inflammatory infiltrates.Altogether, our data demonstrate that LECT2 acts as a strong regulator of liver tumor aggressiveness through its dual action on hepatocytes and impact on the function of tumor infiltrating myeloid cells. This work identifies LECT2 as a new biomarker for HCC and pave the way to new therapeutic strategies
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Ott, Marie-Odile. "Etude de la specificite cellulaire d'expression du gene de l'albumine de rat dans des clones d'hepatome." Paris 7, 1988. http://www.theses.fr/1988PA077131.
Full textAbstract:
Les sequences cis-regulatrices du gene de l'albumine de rat ont ete analysees par transfection transitoire de clones cellulaires d'hepatome de rat presentant differents phenotypes quant a l'expression de l'albumine, mais de meme origine histogenetique. Un fragment de 150 paires de bases en amont du site d'initiation de la transcription suffit a specifier l'expression du gene indicateur et cela uniquement dans les cellules dont le propre gene albumine est exprime. L'extinction selective de l'expression de l'albumine dans des hybrides entre cellules d'un clone differencie d'hepatome de rat et microcellule de fibroblaste de souris depend de la presence d'un chromosome specifique de souris dit "extincteur" de l'albumine. Cette extinction est imposee au promoteur specifique du gene albumine introduit dans les hybrides. Ce promoteur contient donc la (ou les) cible(s) d'une regulatrice
25
Orfanoudakis, Georges. "Diadenosine tetraphosphate : implication dans l'activite mitotique, la replication et la reparation du dna." Université Louis Pasteur (Strasbourg) (1971-2008), 1988. http://www.theses.fr/1988STR13121.
Full textAbstract:
Le diadenosine tetraphosphate (ap**(4)a) est le principal produit de la reaction d'aminoacylation catalysee par certaines aminoacyl-trna synthetases. L'ap**(4)a est une molecule "signal" s'accumulant a l'interphase g1/5 du cycle cellulaire des cellules eucarydes declenchant ainsi la synthese du dna precedant la division cellulaire. Quantification du contenu cellulaire en ap**(4)a et en atp apres synchronisation des cellules (hepertome de rat, fibroblaste de souris) en culture par l'aphidicoline agent bloquant les cellules en phase s et par privation de serum qui arrete la croissance en mi-phase g. Mise au point d'un modele de reparation, dans les ovocytes ou les oeufs non fecondes de xenopus laevis, du plasmute pbr322 modifie par l'acetylaminofluorene. L'effet de l'ap**(4)a sur la reparation est etudie
26
Chi, Chen Hung, and 陳虹琦. "Down-Regulation of p53 in Apoptotic c-Myc-Overexpressing Mouse Mammary Carcinoma Cells." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/51572680473001873574.
Full textAbstract:
碩士國防醫學院
生物化學研究所
91
ABSTRACT The proto-oncogene c-myc plays a crucial role in the proliferation, apoptosis, and regulation of the cell cycle. Aberrant expression of c-Myc could cause apoptosis, but the detail mechanism of c-Myc-induced apoptosis is still not clear. Myc83 cell line has been established from mammary gland tumors of c-Myc-transgenic mice. In the absence of EGF signaling, a lot of Myc83 cells （50％） undergo apoptosis；in the presence of EGF signaling, only 1.5％ Myc83 cells undergo apoptosis. Therefore, we utilize Myc83 cells to study the mechanism of c-Myc-induced apoptosis. Since anisomycin, an activator of p38 MAPK （mitogen-activated protein kinase）and JNK（c-Jun N-terminal kinase）, can induce apoptosis in Myc83 cells, we also utilize anisomycin treated Myc83 cells as another control. We conclude that caspase-3 is activated, PARP is cleaved, the phosphorylation of ERK1 and ERK2 is inhibited in c-Myc-induced apoptosis. In addition, p53 tumor suppressor gene can be regulated by MDM2. MDM2 inhibits the inactivated p53 protein by binding to p53. The decrease of p53 protein and the increase of MDM2 protein were observed in c-Myc-induced apoptosis.p53 mRNA and MDM2 mRNA both increased in c-Myc-induced apoptosis. We propose that c-Myc induced apoptosis is p53-independent.
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Meng, Hsien-Chen, and 孟憲蓁. "The relationship between EMT and drug resistance in c-Myc-overexpressing mouse mammary carcinoma cells." Thesis, 2012. http://ndltd.ncl.edu.tw/handle/03978421973781620862.
Full textAbstract:
碩士國防醫學院
生物化學研究所
100
The epithelial-mesenchymal transition (EMT) is a key mechanism that is often activated during cancer invasion and metastasis. EMT is a multistep process in which cells acquire cellular alterations such as loss of cell-cell junctions and restructuring of the cytoskeleton. In cancer therapy, drug resistance limits the effect of chemotherapy in which cancer cells after simultaneously possess intrinsic or acquired cross-resistance to diverse chemotherapeutic agents.In general, Tumor cells that initially respond to chemotherapy can gradually become resistant and even to different classes of chemotherapeutical drugs resulting in multidrug resistance. Development of multidrug resistance is also the leading cause of cancer recurrence and makes cancer cells behave more aggressive. Recent reports have indicated that the EMT of tumor cells not only increases metastatic potential, but also contributes to drug resistance. We treated Myc83, a c-myc-overexpressing mouse mammary carcinoma cell line, with EGFR tyrosine kinase inhibitor (PD153035) for 22 times to select a drug resistant cell line, DR22. DR22 exhibited more aggregative pattern than parental Myc83. Furthermore, DR22 has shown great potentials of proliferation and soft-agar colony formation. Interestingly, DR22 displayed a downregulation of epithelial and mesenchymal markers and weak motility. Therefore, DR22 exhibits partial EMT phenotype. In addition, a significant reduction of EGFR expression is observed in DR22. We also found that c-Met plays a key role to gain drug resistance in DR22. We characterized the level of EMT mediators and found slug is down-regulated but snail is up-regulated in DR22. Transfection of slug restores the expression of mesenchymal markers. While cross reacting with other anti-cancer drugs, we found that DR22 is resistant to PD153035 only. We therefore hypothesized that there is a close relationship between a complete EMT and multidrug resistance. Thus we treated DR22 with different chemotherapeutic agents (paclitaxel and doxorubicin), and obtained multidrug resistance cells：Taxol 10 and Doxo 20. Consistent with our hypothesis, the multidrug resistant cells display fibroblast phenotype and restoration of mesenchymal markers while E-cadherin remained lost. In addition, we also characterized the level of EMT mediators and found snail was upregulated in Taxol 10 and Doxo 20. Thus, we thought snail may force partial EMT cell (DR22) to undergo complete EMT and exhibits mesenchymal phenotype. Then, we also found c-Met may play a key role to gain multidrug resistance, but we still need more result to prove our hypothesis. Our results elucidates that the down-regulation of EGFR and the increased phosphorylation of c-Met may trigger the resistance to EGFR inhibitor. These phenomenon can also be found in multidrug resistance cells . We also elucidates EMT mediators may play different roles in the progression of EMT program and contribute the process from partial EMT cell to complete EMT.
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Huang, Mei-Chi, and 黃玫琪. "The Signaling Pathway(s) of Drug Resistance in c-Myc-overexpressing Mouse Mammary Carcinoma Cells." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/92109755273278712441.
Full textAbstract:
碩士國防醫學院
生物化學研究所
96
The imbalance among cell proliferation, cell differentiation and apoptosis is implied in tumorigenesis. c-myc, a proto-oncogene, encodes a transcription factor to accelerate cell proliferation, inhibit differentiation and induce apoptosis. Since c-Myc plays crucial roles in cells, the expression of c-Myc is well regulated with short half-life. It has been found that deregulated expression of c-Myc is common in many different cancers and overexpression of c-Myc in cancer usually comes with poor prognosis. Drug resistance in tumor cells could be observed in cancer patients after the treatment of chemotherapy. So far, it is well known that the drug resistant mechanism is induced by multiple drug resistant (MDR). MDR is a transporter of ABC superfamily which can pump the chemotherapy drugs out, however MDR cannot explain the mechanism of resistance to ionic radiation. Myc83 cell line is derived from a tumor of MMTV-c-Myc transgenic mice, thus Myc83 is a c-Myc-overexpressing mouse mammary carcinoma cell line. We utilize Myc83 cell line to study the drug resistant mechanism in c-Myc overexpression condition. We treated Myc83 cell line with EGFR inhibitor PD153035 to develop drug resistance cells (PD cell lines). The drug resistant cells show dramatic resistance to PD153035 with decreased apoptosis ratio and increased proliferation rate. The results show that MDR is similar in parental cells and drug resistant cells, thus MDR could be not involved in the drug resistant mechanism. p53 level is similar during the development of drug resistance, therefore p53 is irrelevant in this drug resistance mechanism. c-Myc level is higher in drug resistant cells than in parental cells; this resalt is consistent with the increased ratio of proliferation. Bax is similar in parental cells and drug resistant cells, thus Bax is also not involved in the drug resistant mechanism. In drug resistant cells, the phosphorylation of p38 MAPK is reduced, which is consistent with the decreased ratio of apoptosis. We also observed the down-regulation of EGFR in drug resistant cells, thus survival signaling cannot be triggered by EGFR and there could be another receptor to trigger the survival signaling. After the treatment of PD153035, we observe the activation of ERK1/2 and Akt are increased, the activation of ERK1/2 and Akt could be one of the major reasons of survival in drug resistant cells when cells were treated with PD153035. PKC is still activated in drug resistant cells, the activation PKC could be another reason of survival in drug resistant cells. p21 level is higher in drug resistant cells than in parental cells, but the ratio of proliferation is higher in drug resistant cells than in parental cells. p21 is not functional in drug resistant cells. Loss of E-cadherin has been seen in resistance cells and this could be involved in epithelial-to-mesenchymal transition (EMT). The character of EMT in cancer cells could play important roles in cell invasion, resistance to anoikis, and angiogenesis. The future work is to confirm whether the drug resistant cells are resistant to other chemotherapy reagents and radiation. Finally, we would like to find the key molecule(s) that may trigger the drug resistance in cancer and build in vivo experiment model to verify.
29
Yu, Ming-Chih, and 游銘志. "The relationship between EMT and drug resistance in c-Myc-overexpressing mouse mammary carcinoma cells." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/74803923390343225604.
Full textAbstract:
碩士國防醫學院
生物化學研究所
98
It is generally known that the proto-oncogene c-MYC , encodes a transcription factor, is implicated in various cellular processes: proliferation, loss of differentiation and apoptosis. In addition, the overexpression of MYC has been observed within a multitude of human tumors, and ranging from hematological malignancies. Previously Myc83, a c-Myc-overexpressing mouse mammary carcinoma cell line, has been treated with EGFR inhibitor (PD153035) to develop a drug resistant cell line, DR22.The loss of E-cadherin and EGFR in DR22 is consistent with the marker of epithelial-mesenchymal transition (EMT) and the morphological change as well. It has been reported that EMT contributes to drug resistance, increasing cell motility and invasion. Therefore, this research focused on characterizing DR22 and the possible involvement of EMT fin cancer progression. Furthermore, the DR22 cells show a specific resistance against PD153035 only but no multiple drug resistance. SB203580 and/or SP600125 partially restored the viability of Myc83 cells under the treatment of different chemotherapy drug. Only SP600125 can partially restore the viability of DR22 cells in doxorubicin. However, the DR22 cells show non-resistance to irradiation, but M phase arrest is increased in radiation treatment. As the results of the wound healing assay and colony assay, we found cell mobility of the DR22 cell was significantly reduced but the tumorigenecity was increased. Since the expression of EMT marker proteins is not consistent, we still observe dramatic down-regulation of E-cadherin. Therefore we suggest that the DR22 display partial EMT. DR22 is more sensitive to other anticancer drugs than parental Myc83 cells. We therefore hypothesized that cells exhibit complete EMT will gain multidrug resistance. Thus we treated DR22 with different chemotherapeutic agents (paclitaxel and doxorubicin), and obtained multidrug resistance cells. Consistent with our hypothesis, the multidrug resistant cells display fibroblastoid phenotype and restoration of mesenchymal markers while E-cadherin and EGFR remained lost. The main purpose of this study is to demonstrate the parallel relationship between the progression from single to multidrug resistance and the development of EMT.
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Abbey, Deepti. "Insights Into Molecular Regulation Of Cardiomyocyte Differentiation Of Mouse Pluripotent Stem Cells." Thesis, 2012. http://hdl.handle.net/2005/2455.
Full textAbstract:
Pluripotent stem cells (PSCs) are specialized cells, which have remarkable ability to maintain in an undifferentiated state and are capable of undergoing differentiation to three germ-layer lineage cell types, under differentiation-enabling conditions. PSCs include embryonic stem (ES)-cells, embryonal carcinoma (EC)-cells and embryonic germ (EG)-cells. ES-cells are derived from the inner cell mass (ICM) of day 3.5 blastocysts (mouse). On the other hand, EC- and EG-cells have different source of origin and exhibit some differences in terms of their differentiation abilities and culture requirements. These PSCs act as an ideal in-vitro model system to study early mammalian development and cell differentiation and, they could potentially be used for experimental cell-based therapy for a number of diseases. However, one of the problems encountered is the immune rejection of transplanted cells. For this, immune-matched induced pluripotent stem (iPS)-cells have been derived from somatic cells, by forced expression of a few stemness genes. Although, human PSCs lines are being experimented, their cell-therapeutic potential is still far from being thoroughly tested due to lack of our understanding regarding lineage-specific differentiation, homing and structural-functional integration of differentiated cell types in the host environment. To understand these mechanisms, it is desirable to have fluorescently-marked PSCs and their differentiated cell-types, which could facilitate experimental cell transplantation studies.
In this regard, our laboratory has earlier generated enhanced green fluorescent protein (EGFP)-expressing FVB/N transgenic ‘green’ mouse: GU-3 line (Devgan et al., 2003). This transgenic mouse has been an excellent source of intrinsically green fluorescent cell types. Recently, we have derived a ‘GS-2’ ES-cell line from the GU-3 mouse line (Singh et al., 2012). Additionally, we envisaged the need for developing an iPS-cell line from the GU-3 mouse and then use them for studying cell differentiation. Thus, aims of the study described in the thesis are to: (1) develop an experimental system to derive EGFP-expressing fluorescently-marked iPS-cell line from a genetically non-permissive FVB/N mouse strain, characterize the established iPS-cell line and achieve differentiation of various cell types from EGFP-expressing iPS-cell line; (2) to study differentiation phenomenon, in particular to cardiac lineage, using select-cardiogenesis
modulators and (3) to assess the gene-expression profiles and signaling system associated with cardiomyocyte differentiation of PSCs.
This thesis is divided into four chapters with the 1st chapter being a review of literature followed by three data chapters. In the chapter I of the thesis, a comprehensive up-to¬date review of literature is provided pertaining to PSCs, their classification, derivation strategies especially for reprogramming of somatic cells for iPSC generation, their differentiation potential and characterization, particularly to cardiac lineage. Various molecular regulators involved in cardiac differentiation of PSCs with emphasis on epigenetic regulation involving DNA methylation and signaling pathways involved are described in detail. Subsequently, various approaches used for enhanced cardiac differentiation of PSCs and the therapeutic potential of PSC-derived differentiated cell types to treat disease(s) are discussed.
Chapter-II describes the successful establishment of a permanent iPS-cell line (named ‘N9’ iPS-cell line) from the non-permissive FVB/N EGFP-transgenic GU-3 ‘green’ mouse. This chapter provides results pertaining to detailed derivation strategy and characterization of the ‘N9’ iPS-cell line which includes colony morphology, expansion (proliferation) efficiency, alkaline phosphatase staining, pluripotent markers’ expression analysis by qPCR and immunostaining approaches and karyotyping analysis. Further, in order to thoroughly assess the differentiation competence of the ‘N9’ iPS¬cell line, assessment of in-vitro and in-vivo differentiation potential of the ‘N9’ iPS-cell line by embryoid body (EB) formation and teratoma formation in nude mice and its detailed histological analysis showing three germ layer cell types and their derivatives were performed, followed by the generation of chimeric blastocysts by aggregation method. This established N9 iPS-cell line could potentially offer a suitable model system to study cardiac differentiation along with other established PSC lines such as the GS-2 and D3 ES-cell lines and the P19 EC-cell line.
Following the establishment of the system to study cardiac differentiation of PSC lines, efforts were made to understand the biology of cardiac differentiation of PSCs (wild¬type and EGFP-transgenic PSC lines and P19 EC-cell line) using small molecules as
modulators. Data pertaining to this is described in Chapter-III. The possible involvement of epigenetic regulation of cardiogenesis for example, DNA methylation changes in cardiogenesis-associated genes is studied using 5-aza cytidine as one of the chromatin modifiers. In order to understand the cardiac differentiation phenomenon, as a consequence of using 5-aza cytidine in cell culture, it was important to investigate its ability to induce/mediate cardiac differentiation. This involved an assessment by quantitating the cardiac beating phenotype and correlating this with enhanced cardiac-gene expression profiles. Further, DNA methylation regulation of cardiogenesis¬associated genes is described using various DNA methylation analysis techniques. Moreover, the possible involvement of other signaling members in mediating the cardiac differentiation is also studied using the P19 EC-cells. Results pertaining to the above findings are described in detail in the Chapter-III.
Chapter-IV is focused on various efforts made towards investigating the ability of ascorbic acid to enhance cardiac differentiation of mouse ES-cells (GS-2 and D3 lines). Ascorbic acid has been implicated to be influencing cardiogenesis and it is reported to enhance differentiation of various cell types under certain culture conditions. Results pertaining to enhancement of cardiac differentiation of PSCs using ascorbic acid are presented in this chapter. This included assessment by quantitating cardiac beating phenotype and its correlation with enhanced cardiogenesis-associated gene expression profiles. Besides, estimation on the sorted cardiomyocyte population, derived from PSCs was also made using mature-cardiac marker. The possible underlying signaling mechanism involved was also studied in detail, using specific inhibitors for pERK (U0126), integrin signaling (pFAK; PP2) and collagen synthesis (DHP), in order to ascertain their involvement in ascorbic acid-mediated cardiac differentiation of mouse ES-cells. Subsequent to the three data chapters (II-IV), separate sections are provided for ‘Summary and Conclusion’ and for ‘Bibliography’, cited in the thesis. The overall scope of the study has been to understand the basic biology of cardiac differentiation from PSCs (EC-cells, iPS-cells and transgenic and wild-type ES-cells) and to assess, by using certain small molecules, whether PSCs could be coaxed to enhance the differentiation to a particular cell type (cardiac). The data contained in this thesis addresses the above theme.
31
Chiou, Liyu, and 邱莉喻. "Studies on chemopreventive potential of garcinol in mouse colon carcinogenesis and inducing growth inhibition in human prostatic carcinoma cells." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/23662795360802775026.
Full textAbstract:
碩士國立高雄海洋科技大學
水產食品科學研究所
99
Garcinol is the polyisoprenylated benzophenone derivative isolated from fruit rind of Garcina indica tropical region. Previous studies demonstrated garcinol possess anti-inflammatory, anti-cancer activities, and chemopreventive efficacy on colonic aberrant crypt foci in rat. However, the effect and mechanisms of garcinol against colitis-associated colon carcinogenesis is yet to be investigated. In this study, we evaluated chemopreventive effect of dietary garcinol on colitis-induced colon carcinogenesis in mice. Six-week ICR mice were injected i.p. with 10 mg/kg azoxymethane (AOM). After one week, 2 % dextran sodium sulfate (DSS) was administered in the drinking for seven days, and then changes ordinary water for following for twenty-five weeks. Our results shown that feeding garcinol (250 ppm and 500 ppm) for twenty-five weeks significantly reduced the tumor size and incidence in mouse colon. Dietary garcinol (500 ppm) also significantly abated colon length induced by AOM/DSS. In addition, western bolt analysis showed that garcinol markedly increased cleavage of PARP, a marker of apoptosis, as well as down-regulated cyclooxygenase-2 (COX-2), proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF) and b-catenin in AOM/DSS-induced colon carcinogenesis in mice. Our results suggested that garcinol might be as potential a chemopreventive agent for colitis-associated colon cancer.
32
chou, chein-Ming, and 周建明. "Study of the influence of p38 MAPK and Mdm2 on p53 degradation in Apoptotic c-Myc-Overexpressing Mouse Mammary Carcinoma Cells." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/85479317631878685678.
Full textAbstract:
碩士國防醫學院
生物化學研究所
92
Myc83 cell line has been established from mammary gland tumors of c-Myc-transgenic mice. PD153035 (an inhibitor of EGFR tyrosine kinase) and Anisomycin (an activator of p38 MAPK and JNK) can induce the down-regulation of p53 in Myc83 cells. This thesis is to focus on the mechanism of p53 down-regulation in c-Myc-induced apoptosis. We utilized SB203580 (an inhibitor of p38 MAPK) and SP600125 (a selective JNK inhibitor) to study the possible signaling pathway and then learned that only SB203580 could inhibit the down-regulation of p53. It means that c-Myc induced down-regulation of p53 through p38 MAPK in Myc83 cells. Furthermore, Mdm2 could bind p53 to induce p53 ubiquitination, transportation from nucleus to cytosol, and further induce degradation of p53 through proteasome. So we utilized MG132 to know the role of Mdm2 in the c-Myc-induced apoptosis and Anisomycin-induced apoptosis. We found that when MG132 was 1μM, it could inhibit proteasome but does not induce aberrant p38 MAPK and apoptosis .We found PD153035 can induce the down-regulation of p53 through Mdm2 but Anisomycin utilized different mechanism. It is well known that radiation can induce p53 expression. Therefore we compare these different apoptosis mechanisms and found that Bcl-xL is induced, caspase-3 is activated, PARP is cleaved in c-Myc-induced apoptosis. Caspase-3 is activated, PARP is cleaved in Anisomycin-induced apoptosis. And radiation-induced apoptosis can activate Caspase-3, PARP.
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Chou, Hua-Kang, and 周華康. "1. Bcl-2 accelerates retinoic acid-induced growth arrest and recovery in human gastric cancer cells. 2. The establishment of animal model and gene therapy in mouse hepatocellular carcinoma." Thesis, 2000. http://ndltd.ncl.edu.tw/handle/60142122900977483615.
Full textAbstract:
博士國防醫學院
醫學科學研究所
88
The role of Bcl-2 as an anti-apoptotic protein has been well documented. In the present work, we present evidence that Bcl-2 may also be involved in cell growth regulation. SC-M1 is an unique cell line which responds to retinoic acid (RA) treatment with reversible growth arrest [Shyu, Jiang, Huang, Chang, Wu, Roffler and Yeh (1995) Eur. J. Cancer 31, 237-243]. In this study, when treated with RA, SC-M1/Bcl2 cells which were generated by transfecting SC-M1 cells with bcl-2 DNA, were growth-arrested two days earlier than SC-M1/neo cells, which were generated by transfecting SC-M1 cells with vector DNA. This indicates that Bcl-2 accelerates RA-induced growth arrest. In addition to the accelerated growth arrest, RA-treated SC-M1/Bcl2 cells also recovered from growth arrest two days faster than SC-M1/neo cells after the removal of RA. Previously, we had identified the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) (p21) as a mediator of RA-induced growth arrest [Tsao, Li, Kuo, Liu and Chen (1996) Biochem. J. 317, 707-711]. In a search for the mechanism by which Bcl-2 affects growth regulation, we found that p21 gene expression was more prominent in SC-M1/Bcl2 cells than in SC-M1/neo cells in the presence of RA; but when RA was removed, p21 gene expression levels in SC-M1/Bcl2 cells were also reduced earlier than SC-M1/neo cells. The present report is the first to show that Bcl-2 accelerates not only growth arrest but also recovery from growth arrest. Moreover, the close correlation between the effect of Bcl-2 on both RA-induced growth arrest and RA-induced p21 gene expression suggests the possibility that Bcl-2 affects cell growth through the mechanism of p21. Hepatocellular carcinoma (HCC) had frequent recurrence problem after primary therapy, then caused poor prognosis. We try to find out a strategy to improve poor prognosis, so utilize the recombinant adenovirus carry the mB7 (Ad-mB7) infect the poor antigenic mouse hepatoma cell line-Hepa1-6 cell. This cell line can express the B7 after FACscan analysis. Hepa1-6 cell inject into the C57BL/6 mouse left lobe liver was performed, then the artificial mouse hepatoma can present through the gross and pathological appearance. In vitro study, the growth curve among the recombinant Ad-mB7 infect hepa1-6 cell (Hepa1-6/Ad-mB7), recombinant Ad-p21 antisense (as) infect hepa1-6 cell (Hepa1-6/Ad-p21 as), and the parental hepa1-6 cell wasn’t different. But in vivo study, Hepa1-6/Ad-mB7 inject into the mouse left lobe liver which show no hepatoma picture. That mean present mB7 could suppress hepatoma formation. Hepa1-6/Ad-mB7 cell inject into CD8 knock-out mouse (KOI) and CD4 knock-out mouse (KOII) left lobe liver respectively, we find out no hepatoma formation in the KOI mouse left lobe liver, but hepatoma formation in the KOII mouse liver appeared. That mean show CD4 is a important role in the suppress hepatoma cell growth using the hepa1-6/Ad-mB7 infect animal model. Hepa1-6/Ad-mB7 and hepa1-6/Ad-p21(as) inject into the C57BL/6 mouse left lobe liver respectively first, then hepa1-6 cell inject into the right subcutaneous skin two weeks later, we find out the hepa1-6/Ad-mB7 the right subcutaneous skin no tumor formation, but the hepa1-6/Ad-p21(as) mouse all died due to the right subcutaneous tumor too large six weeks later. That mean mB7 could prevent hepatoma formation, this is a pilot study in the tumor vaccine experiment.
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Chang, Hung-Chang, and 張宏彰. "The mechanism of drug resistance in c-Myc-overexpressing mouse mammary carcinoma cell line." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/62800776283414344726.
Full textAbstract:
碩士國防醫學院
生物化學研究所
93
The proto-oncogene c-Myc is implicated in various physiological processes: cell growth and apoptosis. Overexpression of c-Myc is associated with many human cancers and further with poor prognosis. In general, acquired drug resistance of tumor cells is frequently observed in cancer patients with the treatment of chemotherapy. Chemotherapy- induced drug resistance is driven by MDR, which could be irrelevant for radiation resistance. However, certain p53 mutants may enhance drug resistance in cancer cells. Myc83 cell line is derived from a tumor of MMTV-c-Myc transgenic mice, thus Myc83 is a c-Myc-overexpressing mouse mammary carcinoma cell line with wild type p53. Myc83 cells were utilized to study the possible relationship between c-Myc and drug resistance. In the current study, we utilized PD153035 (an inhibitor of EGFR tyrosine kinase) and anisomycin (an activator of p38 MAPK and JNK) as useful tools. We found that PD153035 and anisomycin can trigger the down-regulation of p53. We further utilize Myc83 cells to develop drug resistance cells to study the possible mechanism. We hypothesize that p53 has been influenced in Myc83-derived resistant cells. After a serious of treatments, we have selected two independent drug resistant cell clones by repeated treatment with PD153035 and anisomycin. We named these drug resistant cells are PD8 and AN8, respectively. Furthermore, we found these resistant cells are also resistant to IR and 5-FU. In this study, we focus on PD8 cells. Therefore, we tried to find the resistant mechanism of PD8. Our results have demonstrated that: 1. The expression of p53 is not dramatic altered in PD8 cell. 2. In the process to obtain resistant cells, we found the decrease of p53 degradation and the decrease of p38 MAPK activation along with the decrease of apoptosis. Therefore p53 and p38 MAPK could play important roles in the process. 3. When PD8 cells were treated with PD153035, we found the phosphorylation of p38 MAPK is decreased, the degradation of p53 is decreased, and the phosphorylation of Akt and Erk1/2 is increased. Therefore Akt and Erk1/2 could be important in the drug resistant mechanism of PD8. 4. When PD8 cells were treated with 5-FU, the phosphorylation of p38 MAPK is decreased. 5. When PD8 cells were treated with 5-FU and IR, p53 is highly phosphorylated at serine-15. 6. When PD8 cells were treated with MG132 (proteasome inhibitor), p53 degradation is decreased. Our results have revealed some possible factors in drug resistant mechanism, however further experiments need to be performed.
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"Functional characterization of BRE in cell line and chemically-induced mouse liver cancer." 2008. http://library.cuhk.edu.hk/record=b5893556.
Full textAbstract:
Chen, Shuyan.Thesis (M.Phil.)--Chinese University of Hong Kong, 2008.
Includes bibliographical references (leaves 91-98).
Abstracts in English and Chinese.
ABSTRACT --- p.i
ACKNOWLEGGMENTS --- p.v
LIST OF FIGURES --- p.vi
LIST OF TABLES --- p.vii
ABBREVIATIONS --- p.viii
CONTENTS --- p.ix
Chapter Chapter I --- Introduction
Chapter 1.1 --- Introduction of BRE
Chapter 1.1.1 --- Discovery of BRE --- p.1
Chapter 1.1.2 --- Isoforms of BRE --- p.2
Chapter 1.1.3 --- Homology and orthologs of BRE --- p.3
Chapter 1.1.4 --- Expression studies of BRE mRNA --- p.4
Chapter 1.1.5 --- Expression and cellular localization of BRE protein --- p.5
Chapter 1.1.6 --- Interaction between BRE and death receptor --- p.6
Chapter 1.1.7 --- Anti-apoptotic effect of BRE in cell line studies --- p.9
Chapter 1.1.8 --- Anti-apoptotic effect of BRE in vivo --- p.11
Chapter 1.1.9 --- BRE's role in DNA repair and ubiquitination --- p.12
Chapter 1.1.10 --- BRE's role in regulation of Prohibitin and p53 expression --- p.13
Chapter 1.2 --- Hepatocellular carcinoma
Chapter 1.2.1 --- Carcinogenesis --- p.15
Chapter 1.2.2 --- Diethylnitrosamine -induced HCC --- p.15
Chapter 1.2.3 --- Mouse model for HCC studies --- p.17
Chapter 1.2.4 --- BRE in human HCC --- p.18
Chapter 1.3 --- Green Fluorescent Protein
Chapter 1.3.1 --- Application of GFP in biological research --- p.19
Chapter 1.3.2 --- Advantage of GFP applied in protein localization --- p.19
Chapter Chapter II --- Materials and Methods
Chapter 2.1 --- Materials
Chapter 2.1.1 --- Primer used for cloning --- p.20
Chapter 2.1.2 --- DNA clones used in the studies --- p.21
Chapter 2.1.3 --- Materials for DNA manipulation --- p.24
Chapter 2.1.4 --- Materials for protein manipulation --- p.24
Chapter 2.1.5 --- Antibodies --- p.25
Chapter 2.1.6 --- Chemicals --- p.25
Chapter 2.1.7 --- Kits --- p.26
Chapter 2.1.8 --- Culture media and reagents --- p.26
Chapter 2.1.9 --- Bacterial strain used for transformation and cloning --- p.26
Chapter 2.1.10 --- Instrumentation --- p.27
Chapter 2.1.11 --- Animals --- p.27
Chapter 2.1.12 --- Slides --- p.27
Chapter 2.2 --- Methods
Chapter 2.2.1 --- Construction of Plasmids
Chapter 2.2.1.1 --- Polymerase chain reaction (PCR) --- p.28
Chapter 2.2.1.2 --- Enzyme Digestion and Ligation --- p.29
Chapter 2.2.1.3 --- Transformaion
Chapter 2.2.1.3.1 --- Preparation of competent cells --- p.29
Chapter 2.2.1.3.2 --- Heat-shock Transformation --- p.29
Chapter 2.2.1.4 --- Midi Prep of plasmids --- p.30
Chapter 2.2.2 --- Cell Culture --- p.30
Chapter 2.2.3 --- Transfection --- p.30
Chapter 2.2.4 --- MG-132 treatment --- p.31
Chapter 2.2.5 --- Flow Cytometry --- p.32
Chapter 2.2.6 --- Western blotting
Chapter 2.2.6.1 --- SDS-PAGE --- p.32
Chapter 2.2.6.2 --- Immunoblotting --- p.32
Chapter 2.2.7 --- Production of Monoclonal Antibody --- p.33
Chapter 2.2.8 --- Mice --- p.34
Chapter 2.2.9 --- Tissue Processing --- p.35
Chapter 2.2.10 --- Tissue Section --- p.35
Chapter 2.2.11 --- Immunostaining --- p.36
Chapter 2.2.12 --- H&E staining --- p.36
Chapter 2.2.13 --- Picture Capture --- p.37
Chapter 2.2.14 --- Confocal imaging --- p.37
Chapter 2.2.14 --- Statistical Analysis --- p.37
Chapter Chapter III --- BRE promotes growth of chemically-induced hepatocellular carcinoma
Chapter 3.1 --- DEN induced HCC in male mice --- p.38
Chapter 3.2 --- BRE facilitates HCC in female mice --- p.44
Chapter 3.3 --- Over-expression of BRE in tumor portion --- p.45
Chapter 3.4 --- Direct effect of DEN on BRE expression --- p.47
Chapter 3.5 --- Contribution of infiltrating cells in up-regulation of BRE --- p.50
Chapter Chaper IV --- Subcellular localization of BRE
Chapter 4.1 --- GFP-BRE fusion constructs --- p.55
Chapter 4.1.1 --- Transfection of GFP-BRE fusions --- p.58
Chapter 4.1.2 --- Flow cytometry analysis of GFP-BRE fusions --- p.59
Chapter 4.1.3 --- Western blot analysis of GFP-BRE fusions --- p.62
Chapter 4.1.4 --- Stabilities of GFP-BRE fusions --- p.64
Chapter 4.2 --- Fusions between GFP and the deletion mutants of BRE --- p.66
Chapter 4.2.1 --- Transfection of mutants --- p.68
Chapter 4.2.2 --- Low expression of mutants --- p.69
Chapter 4.3 --- MG-132 treatments
Chapter 4.3.1 --- Increased expression of fusion proteins --- p.74
Chapter 4.3.2 --- Subcellular localization of GFP-BRE fusions --- p.77
Chapter Chapter V --- Discussion
Chapter 5.1 --- Functional role of BRE in HCC
Chapter 5.1.1 --- Stage model of carcinogenesis --- p.81
Chapter 5.1.2 --- Anti-apoptotic genes in cancer --- p.84
Chapter 5.1.3 --- Limitation of the study --- p.85
Chapter 5.1.4 --- Conclusion --- p.85
Chapter 5.2 --- Subcellular localization of BRE
Chapter 5.2.1 --- Low expression of GFP-BRE fusions --- p.86
Chapter 5.2.2 --- Additional study --- p.90
Chapter 5.2.3 --- Conclusion --- p.90
Reference --- p.91
Appendix --- p.99
36
Li, Hsin-Ying, and 李欣穎. "p53 Down-regulation May Contribute to Drug Resistance in c-Myc-Overexpressing Mouse Mammary Carcinoma Cell Line." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/49082632223727521838.
Full textAbstract:
碩士國防醫學院
生物化學研究所
94
The proto-oncogene c-Myc is implicated in different physiological processes: cell growth and apoptosis. Overexpression of c-Myc is associated with many human cancers and further with poor prognosis, however the mechanism of c-Myc induced apoptosis is not clear. In general, acquired drug resistance of tumor cells is frequently observed in cancer patients with the treatment of chemotherapy. Chemotherapy- induced drug resistance is driven by MDR, which could be irrelevant for radiation resistance. However, certain p53 deregulated may enhance drug resistance in cancer cells. Myc83 cell line is derived from a tumor of MMTV-c-Myc transgenic mice, thus Myc83 is a c-Myc-overexpressing mouse mammary carcinoma cell line with wild type p53. Myc83 cells were utilized to study the protein participate in c-Myc-induced apoptosis and the possible relationship between c-Myc and drug resistance. In the current study, we utilized PD153035 (an inhibitor of EGFR tyrosine kinase) and anisomycin (an activator of p38 MAPK and JNK) as useful tools. In the absence of survival signaling driven by EGFR, Myc83 cells undergo c-Myc-induced apoptosis. We have found that both PD153035 and anisomycin can trigger the down-regulation of p53. We further utilize Myc83 cells to develop drug resistance cells to study the possible mechanism. We prolong the time of treatment from 24 hoursto 48 hours to let the cell growth in more sever enviroment, and avoid the phenomenon of drugs resistant is simply caused by the delay of apoptosis. Subsequently we get the super drug resistant cells. We subculture the super drug resistant cells and let the cell fully respond to the damage. Finally we find the level of p53 is decreased in the late passage of the super drugs resistant cells. We also find the activation of p38MAPK is elveated in the late passage of the drugs resistant cells. To further confirm our result we also use a general chemotherapy drug 5-Flurouracil as treatment and obtain the drugs resistant cells named M5-1, M5-2 and M5-3. The activation of p38MAPK is also increased in the drugs resistant cell selected by 5-Flurouracil. Therefore we demonstrated that p38MAPK may induce drugs resistance in c-Myc overexpressing cells.
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CHUN-CHIEH, CHANG, and 張俊傑. "Epidermal growth factor receptor inhibitor (EGFRI)-induced drug resistance in c-Myc-overexpressing mouse mammary carcinoma cell line." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/37096228890649163888.
Full textAbstract:
碩士國防醫學院
生物化學研究所
95
In general, acquired drug resistance is frequently observed in cancer patients with the treatment of chemotherapy. Chemotherapy-induced drug resistance is driven by MDR, which could be irrelevant for radiation resistance. Therefore﹑we hyphothesize there should be some other factor(s) contribute to drug resistance. According to Epidemiology, overexpression of c-Myc is associated with poor prognosis. Thus we utilized Myc83,a c-Myc-overexpressing mouse mammary carcinoma cell line, to study the possible relationship between c-Myc and drug resistance. Myc83 cells were treated with PD153035 to develop drug resistance cells ( PD cell line ).To obtain stronger drug-resistant cell clones , we prolong the time of treatment from previous 24 hours to 48 hours and avoid the possibility that the drug resistantance is caused by the delay of apoptosis. After a serial of treatments, we have selected different independent drug resistant cell clones by repeating treatment of PD153035. We named these drug resistant cells：PD8、PD14 and PD22,respectively.It is consisitant with MDR related papers that these resistant cells are also resistant to IR and 5-FU.To learn the drug resistance mechanism, we examined the morphology and proliferation rate and found dramatic change along with the increment of the PD153035 treatment.Oru previous results have shoen that PD153035 can trigger the down-regulation of p53, there we wanted to observe the level of p53 and subcultured the drug resistant cells and lets the cell fully respond to the impact of PD153035. Interestingly we find the level of p53 is decreased after individual drug resistant cell line subculture. However, p53 level is reversed in the late passage. Since p53 is critical for cancer therapy, we believe that there could be some relationships between drug-resistantance and the downregulation of p53. we also found the phosphorylation of p53 (serine-15) is decreased in early passage,but increased in late passage. when we checlced some signaling pathways, we also found the phosphorylation of p38 MAPK, and Akt, and Erk1/2 are increased. Therefore p53, p38 MAPK, Akt and Erk1/2 could be important in c-Myc-induved drug resistant mechanism. Our results have revealed some possible factors in c-Myc induced drug resistant mechanism, however further experiments need to be performed.
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Linder, Benedikt. "Interaction of the Hedgehog and vitamin D receptor signaling pathways in Patched associated cancers." Doctoral thesis, 2015. http://hdl.handle.net/11858/00-1735-0000-0022-5FDC-A.
Full text