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The evaluation of senescence markers in the H9c2 cell line after cobalt exposure
Rottenbornová, Lucie ; Jirkovský, Eduard (advisor) ; Hrubša, Marcel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Lucie Rottenbornová Supervisor: PharmDr. Eduard Jirkovský, Ph.D. Title of diploma thesis: The evaluation of senescence markers in the H9c2 cell line after cobalt exposure Introduction and aim: Cellular senescence is the response of a cell to a physiological or stressful stimulus. Cobalt is an element that simulates hypoxia under normoxic conditions through hypoxia-inducible factors. One of the factors that can induce cellular senescence is hypoxia. The aim of the work was to evaluate p16INK4a and Ki67 proteins as markers of cell senescence in an H9c2 cell line after exposure to CoCl2. Methods: Cytotoxicity was measured using the MTT assay on the H9c2 cell line after 24 and 48 hours of exposure to CoCl2. Subsequently, relative gene expression of genes associated with hypoxia (HIF1α and BNIP3) and senescence (p16INK4a and Ki67) in H9c2 after exposure to cobalt was investigated using the RT-PCR method. Results: Exposure of H9c2 cells to CoCl2 induced dose- and time-dependent toxicity. After a 24-hour exposure, CoCl2 toxicity was not observed up to a concentration of 100 μM, the IC50 value was 500.5 μM. After 48 hours of exposure, higher toxicity was observed already at 100 μM concentration and...
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Molecular analysis of head and neck carcinomas associated with HPV infection
Glendová, Kristýna ; Staněk, Libor (advisor) ; Tachezy, Ruth (referee)
Head and neck cancers (HNSCC) are highly heterogeneous disease, results from two major carcinogens - tobacco and/or alcohol, or HR HPV infection. This thesis was based on 60 biopsies of head and neck tumours embedded into paraffin after histological verification. HPV infection, including particular types was monitored in different HNSCC regions by multiplex qPCR. Subsequent IHC demonstrated expression of p16INK4A and p53 as a possible diagnostic biomarker. Based on the information, patients with HNSCC can benefit from antiEGFR therapy by Cetuximab, but so far without defined predictors, the analysis of point mutations of Ras gene family (Kras, Nras) and Braf gene was performed. These mutations were monitored as potential predictive biomarkers, in correlation with gender, age and other risk factors. For all statistical processing the Chi-x2 test was used. Key words: Head and neck cancers, biopsy, HPV types, PCR, p16INK4A, p53, molecular predictors, Kras, Nras, Braf
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Molecular analysis of head and neck carcinomas associated with HPV infection
Glendová, Kristýna ; Staněk, Libor (advisor) ; Tachezy, Ruth (referee)
Head and neck cancers (HNSCC) are highly heterogeneous disease, results from two major carcinogens - tobacco and/or alcohol, or HR HPV infection. This thesis was based on 60 biopsies of head and neck tumours embedded into paraffin after histological verification. HPV infection, including particular types was monitored in different HNSCC regions by multiplex qPCR. Subsequent IHC demonstrated expression of p16INK4A and p53 as a possible diagnostic biomarker. Based on the information, patients with HNSCC can benefit from antiEGFR therapy by Cetuximab, but so far without defined predictors, the analysis of point mutations of Ras gene family (Kras, Nras) and Braf gene was performed. These mutations were monitored as potential predictive biomarkers, in correlation with gender, age and other risk factors. For all statistical processing the Chi-x2 test was used. Key words: Head and neck cancers, biopsy, HPV types, PCR, p16INK4A, p53, molecular predictors, Kras, Nras, Braf
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