目的 观察隐丹参酮对人膀胱癌T24及UMUC-3细胞自噬的影响并讨论其可能的机制.方法 通过体外培养T24及UMUC-3细胞,用40、80、120、160 μmol/L隐丹参酮处理,以CCK-8检测各组细胞的增殖抑制率,并计算IC50.采用IC50药物浓度处理T24及UMUC-3细胞后,通过 Western Blot分析检测自噬相关蛋白LC3、Beclin-1和PI3K/Akt/mTOR信号通路蛋白的表达.结果 CCK-8结果提示在一定时间和浓度范围,隐丹参酮浓度与膀胱癌细胞的生长抑制率成正比,各个实验组与对照组相比差异均有统计学意义(P＜0.05).Western Blot分析结果显示,与对照组(NC)比较,隐丹参酮组(cry)T24和UMUC-3细胞LC3-Ⅱ/LC3-I比值明显升高,提示LC3-I向LC3-1I转化增加,同时Beclin-1表达水平明显升高,p-PI3K、p-Akt和p-mTOR蛋白表达水平明显降低,差异有统计学意义(P＜0.01).结论 隐丹参酮可能通过抑制PI3K/Akt/mTOR信号通路的活化,从而诱导膀胱癌T24和UMUC-3细胞发生自噬.
Effects of Cryptotanshinone on Autophagy in T24 and UMUC-3 cells in Bladder Cancer
Objective To observe the effects of cryptotanshinone on autophagy in human bladder cancer T24 and UMUC-3 cells and discuss its possible mechanisms.Methods T24 and UMUC-3 cells were cultured in vitro and treated with 40,80,120 and 160μmol/L cryptotanshinone.The proliferation inhibition rates of the cells were detected by CCK-8,and IC50 was calculated.After T24 and UMUC-3 cells were treated with IC50 concentration,the expressions of autophagy related proteins LC3,Beclin-1 and PI3K/Akt/mTOR signaling pathway proteins were detected by Western Blot analysis.Results The results of CCK-8 indicated that in a certain time and concentration range,the concentration of cryptotanshinone was directly proportional to the growth inhibition rate of bladder cancer cells,and there were statistical differences between each experimental group and the control group(P＜0.05).Western Blot a-nalysis showed that compared with the control group(NC),LC3-Ⅱ/LC3-I ratio of T24 and UMUC-3 cells in cryptotanshinone group(cry)was significantly increased,indicating that LC3-I to LC3-Ⅱ transformation was increased,and Beclin-1 expression level was significantly increased.The protein expression levels of P-PI3K,P-Akt and P-mTOR were significantly decreased,with statistic differences(P＜0.01).Conclusion Cryptotanshinone may induce autophagy in bladder cancer T24 and UMUC-3 cells,possibly by inhibiting the activation of the PI3 K/Akt/mTOR signaling pathway.
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