星形孢菌素通过PI3K/Akt信号途径影响人宫颈癌细胞增殖与侵袭能力的研究

摘要

目的 探讨蛋白激酶C(PKC)抑制剂星形孢菌素(STS)对人宫颈癌细胞增殖与侵袭能力的影响,并初步探讨其信号机制.方法 培养人宫颈癌细胞Hela细胞株,建立STS组与空白对照组.采用MTT法观察各组细胞增殖能力,Transwell检测细胞侵袭能力,以Real-time PCR与Western Blot检测细胞PKCα、磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(Akt)在mRNA水平与蛋白水平表达量以及对PI3K磷酸化的作用.结果 MTT实验中,0.5μmol/L STS组24 h后吸光度A值显著低于空白对照组,而增殖抑制率显著高于空白对照组(P<0.05);Transwell实验STS组穿越细胞数少于空白对照组[(77.3±9.4)个vs.(127.3±6.8)个],差异有统计学意义(P<0.05);Real-time PCR结果显示STS组PKCα、PI3K、Akt mRNA表达量分别为空白对照组的43.4％、65.8％、55.6％,差异有统计学意义(P<0.05);Western Blot检测STS组PKCα、PI3K、Akt、P-PI3K蛋白表达量为空白对照组的34.7％、38.8％、80.6％,75.3％,差异有统计学意义(P<0.05).结论 STS可以有效抑制人宫颈癌细胞Hela的增殖与侵袭能力,其机制可能与降低PKCα的表达后抑制了下游PI3K/Akt信号的激活有关.%Objective To investigate the effect of protein kinase C inhibitor (STS)on the proliferation and invasion of human cervical cancer cells,and to explore its mechanism. Methods Hela cell lines were divided into STS group and control group,MTT method was used to observe the cell proliferation ability,and the invasion ability of cells was detected by Transwell method,the expression of mRNA and protein of PKCα、PI3K、Akt and the effetc of PI3K phosphorylation by Real - time quantitative polymerase chain reaction (Real - time qPCR)and Western Blot. Results Cell proliferation ability and invasion ability of STS group were significantly lower than that of control group (P < 0. 05). The mRNA levels of PKC,PI3K in STS group were 43. 4％,65. 8％ and 55. 6％ respectively to the control group (P < 0. 05). The protein levels of PKC,PI3K,Akt and P - PI3K were 34. 7％,38. 8％,80. 6％,75. 3％ respectively to the control group (P < 0. 05). Conclusion STS can effectively inhibit the proliferation and invasion of human cervical cancer cell line Hela,and its mechanism may be related to the decrease of PKC expression and the activation of downstream PI3K/ Akt signal.