基于wnt/β-catenin通路探讨雷公藤内酯醇调控人卵巢癌SKOV-3细胞迁移机制

Study on the Mechanism of Regulation of Triptolide on SKOV-3 Cells Migration in Human Ovarian Cancer Based on wnt/β-catenin Pathway

目的 探讨雷公藤内酯醇(TPT)对人卵巢癌SKOV-3细胞迁移能力的影响及其通过调控wnt/β-catenin影响SKOV-3细胞迁移的机制。方法 将实验分为5组,空白对照组(不给予任何药物处理)、低浓度TPT组(10 nmol·L^(-1) TPT作用24 h)、中浓度TPT组(50 nmol·L^(-1) TPT作用24 h)、高浓度TPT组(100 nmol·L^(-1) TPT作用24 h)、β-catenin抑制剂组(5μmol·L^(-1) IWR-1-endo作用24 h)。MTT法检测SKOV-3细胞存活情况;Transwell法检测SKOV-3细胞迁移情况;Western Blot法检测SKOV-3细胞蛋白表达;RealtimePCR检测SKOV-3细胞mRNA的表达。结果 与空白对照组相比,低、中、高浓度TPT组SKOV-3细胞存活率、迁移率均逐渐降低,且不同浓度TPT组间的细胞存活率差异具有统计学意义(P<0.05);β-catenin抑制剂组与空白对照比较,SKOV-3细胞存活率、迁移率均显著降低(P<0.05)。与空白对照组相比,低、中、高浓度TPT组wnt、β-catenin、c-Myc、TCF-1、cyclinD1、MMP-7蛋白及mRNA的表达水平均明显降低,且呈一定浓度依赖性(P<0.05);β-catenin抑制剂组wnt、β-catenin、c-Myc、TCF-1、cyclinD1、MMP-7蛋白及mRNA的表达水平均低于空白对照组(P<0.05)。结论 TPT对Wnt/β-catenin信号通路相关蛋白表达具有负向调控作用,进而抑制卵巢癌SKOV-3细胞迁移。

OBJECTIVE To investigate the effects of Tripterygiolide(TPT) on the migration of human ovarian cancer SKOV-3cells and the mechanism of SKOV-3 cells migration by regulating wnt/β-catenin.METHODS The experiment was divided into 5 groups:blank control group(no drug treatment),low concentration TPT group(10 nmol·L~(-1) TPT for 24 h),medium concentration TPT group(50 nmol·L~(-1) TPT for 24 h),high concentration TPT group(100 nmol·L~(-1) TPT for 24 h),β-catenin Inhibitor group(5 μmol·L~(-1) IWR-1-endo for 24 h).The survival of SKOV-3 cells was detected by MTT assay.SKOV-3 cells migration were detected by Transwell method.The protein expression of SKOV-3 cells was detected by Western Blot.The mRNA expression of SKOV-3 cells was detected by RealtimePCR.RESULTS Compared with blank control group,the survival rate and mobility of SKOV-3 cells in low,medium and high concentration TPT groups were decreased gradually,and the survival rate of SKOV-3 cells in different concentrations of TPT groups was statistically different(P<0.05).Compared with blank control,the survival rate and mobility of SKOV-3 cells in β-catenin inhibitor group were significantly decreased(P<0.05).Compared with blank control group,the expression levels of wnt,β-catenin,c-Myc,TCF-1,cyclinD1 and MMP-7 protein and mRNA in low,medium and high concentration TPT groups were significantly decreased in a concentration-dependent manner(P < 0.05).The protein and mRNA expression levels of wnt,β-catenin,c-Myc,TCF-1,cyclinD1 and MMP-7 in β-catenin inhibitor group were lower than those in blank control group(P<0.05).CONCLUSION TPT can negatively regulate the expression of wnt/β-catenin signaling pathway and inhibit SKOV-3 cells migration in ovarian cancer.