【作者】 孙欣；

【导师】 于德新；

【作者基本信息】 安徽医科大学 ， 外科学（泌尿外科学）， 2016， 硕士

【摘要】 目的:联苯胺,属于芳香胺类物质,被认为是已知的与膀胱癌密切相关联的人体致癌物。然而,如今关于联苯胺促进肿瘤进展的研究还是比较匮乏。本研究中主要探讨在联苯胺对人膀胱癌T24细胞的ERK1/2/AP-1通路的激活和细胞周期调控的影响,并确定姜黄素是否能对联苯胺诱导的改变起到化学预防作用。方法:人膀胱癌T24细胞用不同浓度的联苯胺(0-5μΜol/l)处理6天,或与ERK1/2的特异性抑制剂U0126(0-15μΜol/l)或姜黄素(0-20μΜol/l)共同处理6天,MTT法检测细胞活力及用流式细胞仪检测细胞周期。Western blot及RT-PCR分别检测ERK1/2/AP-1通路,Cyclin D1,PCNA和p21的表达。结果:低浓度的联苯胺(0.001,0.005和0.01μΜol/l)可以促进T24细胞的细胞增殖和增强ERK1/2蛋白的磷酸化。我们还发现AP-1蛋白的表达亦增强。此外,联苯胺处理T24细胞引起细胞周期相关蛋白cyclin D1和PCNA的表达上调,以及P21的表达下调。后期我们用联苯胺联合ERK1/2的特异性抑制剂U0126(10μΜol/l)或姜黄素(5μΜol/l)共同处理细胞6天,结果发现ERK1/2的特异性抑制剂U0126及姜黄素可以逆转联苯胺引起的细胞改变。结论:以上研究表明,低浓度联苯胺通过激活ERK1/2信号通路促进人膀胱癌T24细胞的增殖。姜黄素可能为联苯胺相关膀胱癌提供新的治疗策略。更多还原

【Abstract】 Objective: Benzidine, which belongs to aromatic amines, is classified as a known human carcinogen which is closely associated with bladder cancer. However, little knowledge is available concerning its cancer-promoting activities. The present study described here aimed to investigate the effects of benzidine on ERK1/2/AP-1 activation and cell cycle regulation in human bladder carcinoma T24 cells and determine whether curcumin has a protective role in benzidine-induced alteraions. Methods:Exposing T24 cells to various concerntions of benzidine with or without U0126 or curcumin for 6 days, cell viability was evaluated by MTT assay and cell cycle was detected by flow cytometry assay subsequently. Expressions of ERK1/2/AP-1 proteins, cyclin D1, PCNA and p21 were also assessed by western blot and RT-PCR. Results:Low concentrations of benzidine(0.001, 0.005 and 0.01 μΜol/l) promoted cell proliferation and enhanced ERK1/2 phosphorylation in T24 cells. Distinct expressions of AP-1 proteins were also observed. Furthermore, we found that treating T24 cells with benzidine induced the up-regulation of cyclin D1 and PCNA as well as down-regulation of p21. Afterwards, cells were co-treated with U0126(10 μΜol/l) or curcumin(5 μΜol/l) for 6 days. Then we found benzidine-induced alteraions in T24 cells were reversed. Conclusion:These findings indicated that low concentrations of benzidine enhanced cell proliferation in T24 cells. Inhibition of ERK1/2 pathway by U0126 suppressed benzidine-induced cell proliferation. Curcumin may serves as an effective agent against benzidine-associated bladder cancer.更多还原