膀胱癌顺铂耐药与其基因甲基化相关性分析-correlation between cisplatin resistance and gene methylation in bladder cancer.docx

膀胱癌顺铂耐药与其基因甲基化相关性研究英文摘要 膀胱癌顺铂耐药与其基因甲基化相关性研究 英文摘要 PAGE PAGE III IV IV Study about Cisplatin-resistance of bladder cancer cells and their DNA methylation pattern Abstract Objective: To investigate the DNA methylation profile of cisplatin-resistance in bladder cancer cell, and explore the alteration of genes and proteins which are involved in cisplatin-resistance. Methods: The build-up of cisplatin-resistant bladder cancer cells (T24/DDP): The cisplatin-resistant bladder cancer cells were built up in T24 bladder cancer cells (T24/DDP) with continued increasing of concentrations (0.2-2.0mg/L). After the built-up of T24/DDP cell line, a series of biological parameters were measured including cisplatin sensitivity, cellular morphology, growth curve and cell cycle. DNA methylation profile and selection of drug-resistant genes in T24/DDP cell line: The DNA methylation profile was conducted with DNA methylation chips in T24 and T24/DDP cells, and genes with hypermethylation or hypomethylation were screened by statistical methods. The mRNA expression of selected genes with altered methylation was analyzed with real time-PCR. Then, the methylation of these genes was further validated with methylation-specific PCR (MSP). The biological impacts of 5-aza-dC on T24/DDP cells: The demethylation was conducted in T24/DDP cells by 5-aza-dC treatment. Then, a series of cellular biological parameters were measured. The molecular mechanisms of demethylation with 5-aza-dC in T24/DDP cells: The expression of genes and proteins were analyzed with real time-PCR and western blot in T24/DDP cells treated by 5-aza-dC. Results: 英文摘要 膀胱癌顺铂耐药与其基因甲基化相关性研究 The cisplatin-resistant bladder cancer cells (T24/DDP) were successfully built up in T24 cell line. T24 cells were almost 100% died of cisplatin treatment at the concentration of 20 ug/ml, whereas it was only 30% in T24/DDP cells. The doubling time of T24 and T24/DDP cells was 72.5 and 90.2 hours respectively. There was no obvio