p53/miR-34a通路介导羽扇豆醇的抑制人膀胱癌T24细胞增殖作用  

Anti-proliferative Effect of Lupeol on Human Bladder Cancer T24 Cell Line via p53/miR-34a Signaling

作　　者：郭敏[1] 刘佩[2] 郑国华[2] 邱振鹏[2] 

机构地区：[1]华中科技大学同济医学院附属同济医院药学部,武汉430030 [2]湖北中医药大学药学院

出　　处：《中国药师》2016年第9期1629-1632,共4页China Pharmacist

基　　金：国家自然科学基金资助项目(编号:81173502)

摘　　要：目的：探讨羽扇豆醇对人膀胱癌T24细胞增殖的影响及对p53／miR-34a通路调控机制。方法：运用CCK-8法检测不同浓度羽扇豆醇（5—120μmol·L^-1）分别作用24h和48h对T24细胞增殖的影响；运用CCK-8法结合Caspase抑制药确证参与羽扇豆醇诱导细胞死亡的Caspase亚型；分别运用荧光定量PCR（qPCR）和蛋白免疫印记评价羽扇豆醇对miR-34a及p53蛋白表达的影响；运用qPCR评价羽扇豆醇对miR-34a下游靶基因Bcl-2、CD44、c-Myc的mRNA表达的影响。结果：T24细胞经羽扇豆醇处理后，其细胞增殖受到明显抑制，且呈一定的剂量依赖性；药物作用24h和48h时羽扇豆醇的半抑制浓度（IC50）分别为（77．23±6．78），（64．58±4．23）μmol·L^-1。与对照组相比，羽扇豆醇能够使T24细胞中p53蛋白表达上调，还能够增加miR-34a表达水平，差异具有统计学意义（P〈0．01）；羽扇豆醇处理后细胞内Bcl-2、CD44、c—Myc的mRNA表达量下调，差异具有统计学意义（P〈0．01）。结论：羽扇豆醇具有抑制膀胱癌似细胞增殖能力，其作用机制与调控p53／miR-34a通路有关。Objective： To study the anti-proliferative effect of lupeol on human bladder cancer T24 cell line and the regulating mechanism for p53/miR-34a signaling. Methods： CCK-8 assay was performed to evaluate the effects of lupeol at different concentrations on cell viability in 24 h and 48 h. Caspase inhibitors were used to identify the subtypes of Caspase during lupeol induced cell death. The effects of lupeol on the expression of total p53 protein and miR-34a were evaluated by western blot and real-time PCR, respectively. The effects of lupeol on downstream targets of miR-34a were quantified by real-time PCR. Results： Lupeol could inhibit the proliferation of T24 cells in a dose-dependent manner. The IC50 of lupeol was （77.23 ±6.78 ） μmol·L^-1 in 24 h. Compared with the control group, lupeol could elevate the expressions of p53 and miR-34a （P 〈 0.01 ）. Moreover, the mRNA expression of miR-34a targets, Bcl-2, CD44 and c-Myc were significantly down-regulated after the treatment with lupeol （P 〈 0.01 ）. Conclusion： Lupeol can inhibit T24 cell proliferation, which is related with the regulating effects on p53/miR-34a signaling.

关 键 词：羽扇豆醇 p53/miR-34a通路 膀胱癌 

分 类 号：R965.1[医药卫生—药理学]