Hsp90潜在抑制剂姜黄素诱导人宫颈癌Hela细胞凋亡的机制

The mechanisms of apoptosis in Hela cells induced by Hsp90 potential inhibitor Curcumin

目的:研究姜黄素对人宫颈癌细胞Hela的抑制作用及对Hsp90信号通路的影响。方法:MTT法检测姜黄素对Hela细胞的增殖抑制作用。划痕实验检测姜黄素对细胞浸润侵袭的影响。流式细胞术检测姜黄素促进细胞凋亡的效果。Western blotting检测姜黄素对Hela细胞内Hsp90、Hsp70、AKT及Her-2蛋白表达的影响。结果:作用48h后,姜黄素对Hela细胞的增殖抑制呈现出显著的剂量依赖性(P<0.01),当药物浓度为100μmol/L时抑制率达到(87.5±3.5)%,IC50值为(41.8±2.3)μmol/L。划痕实验表明,姜黄素能抑制细胞的浸润和迁移。流式细胞结果显示,姜黄素能有效的促进Hela细胞凋亡,40μmol/L组细胞的凋亡率即从原来的(10.3±0.6)%提高至(19.1±1.5)%(P<0.05),当药物浓度达到100μmol/L时凋亡率达到(55.8±2.6)%。Western blotting结果显示,姜黄素能引起Hsp70蛋白表达上调,并促进Hsp90下游客户蛋白AKT及Her-2降解。结论:姜黄素表现出较强的抗Hela细胞活性,其机制可能是通过抑制Hsp90活性,影响相关信号通路引起的。

Objective：To observe the effect of Curcumin on inhibition of human cervical carcinoma cell lines Hela,and investigate the potential mechanism on Hsp90 signal pathway.Methods：MTT assay was used to detect the anti -proliferative ability of Curcumin on Hela cells.The inhibition effect of curcumin on cell infiltration and invasion was detected by Wound scratch assay.Flow cytometry was used to detect the influence of Curcumin on the apoptosis of Hela cells.The expressions of Hsp90 signal pathway-related proteins such as Hsp90,Hsp70,AKT and Her-2 in Hela cells were determined by Western blotting.Results：After treated with Curcumin for 4 8 h,proliferation of Hela cells was inhibitedby Curcumin in a significant dose-dependent manner（P 〈 0.01）,the IC50 value was （41.8 ± 2.3） μmol/L.When the concentration of Curcumin maintained at 100μmol/L,the inhibition rate was up to （87.5 ± 3.5） %.Wound scratch experiments showed that Curcumin had ability of preventing the invasion and migration of Hela cells.Flow cytometry assay showed that Curcumin can effectively promote apoptosis to Hela cells.Compared with control group（10.3 ±0.6）%,apoptosis rate of Hela cells were arised from （19.1 ± 1.5）% to （55.8 ± 2.6）% as the concentration of Curcumin increased from 40μmol/L to 100μmol/L （P 〈 0.01）.Western blotting experiment showed that Curcumin was able to increase expression of Hsp70,while promoted degradation of AKT and Her-2 which were client proteins of Hsp90.Conclusion：Curcumin is able to inhibit the proliferation of Hela cells and promote it apoptosis,the potential mechanism that probably is its affection on Hsp90-related signaling pathways by inhibiting activity of Hsp90.