雷公藤红素调控Hedgehog通路促进人卵巢癌SKOV-3细胞凋亡

Tripterine promotes apoptosis of human ovarian cancer SKOV-3 cells by regulating Hedgehog pathway

目的探讨雷公藤红素(tripterine)对人卵巢癌SKOV-3细胞凋亡的影响及其通过调控Hedgehog通路影响SKOV-3细胞凋亡的机制。方法将实验分为空白对照组,雷公藤红素低剂量(5nmol/L)组、雷公藤红素中剂量(10nmol/L)组、雷公藤红素高剂量(20 nmol/L)组和环巴胺(cyclopamine)抑制剂(5μmol/L)组5组。MTT检测SKOV-3细胞存活率。流式细胞术检测SKOV-3细胞凋亡,Western blot和Real time PCR检测SKOV-3细胞中SHH、PTCH1、Gli1、Gli3、SMO及下游Bcl-2、Caspase-3的蛋白和mRNA表达水平。结果与空白对照组进行比较,雷公藤红素低、中、高剂量组及环巴胺抑制剂组细胞存活率降低,细胞凋亡率升高,SHH、PTCH1、Gli1、SMO、Bcl-2蛋白及其mRNA相对表达量降低,Gli3、Caspase-3蛋白及mRNA相对表达量升高。与雷公藤红素低剂量组比较,雷公藤红素中、高剂量组及环巴胺抑制剂组细胞存活率降低,细胞凋亡率升高,SHH、PTCH1、Gli1、SMO、Bcl-2蛋白及其mRNA相对表达量降低,Gli3、Caspase-3蛋白及m RNA相对表达量升高,雷公藤红素高剂量组上述指标变化程度更明显。结论雷公藤红素通过调控Hedgehog通路及其下游基因的表达促进人卵巢癌SKOV-3细胞凋亡。

Objective To investigate the effect of tripterine on the apoptosis of human ovarian cancer SKOV-3 cells and its mechanism.Methods The cultured cells were divided into five groups:blank control group,low-dose(5 nmol/L) tripterine group,medium-dose(10 nmol/L) tripterine group,high-dose(20 nmol/L) tripterine group and cyclopamine inhibitor(5μmol/L) group.The survival rate of SKOV-3 cells was detected by MTT,the apoptosis of SKOV-3 cells was detected by flow cytometry,and the protein and mRNA expression level of SHH,PTCH1,Gli1,Gli3,SMO and downstream Bcl-2,Caspase-3 were detected by Western blot and Real-time PCR.Results Compared with the blank control group,the cell survival rate of low,medium and high-dose tripterine groups and cyclopamine inhibitor groups were decreased,while the apoptosis rate were increased,the relative protein and mRNA expression of SHH,PTCH1,Gli1,SMO and Bcl-2 were decreased,and the relative protein and mRNA expression of Gli3 and Caspase-3 were increased.Compared with the low-dose tripterine group,the cell survival rate of the medium and high-dose tripterine groups and cyclopamine inhibitor groups were decreased,the apoptosis rate were increased,the relative protein and mRNA expression of SHH,PTCH1,Gli1,SMO and Bcl-2 were decreased,and the relative protein and mRNA expression of Gli3 and caspase-3 were increased.The highdose group showed the most significant changes among all the groups.Conclusion Tripterine promotes apoptosis of human ovarian cancer SKOV-3 cells by regulating the Hedgehog pathway and its downstream genes.