SHBG 基因在人滋养细胞凋亡中的作用

摘要

目的：研究性激素结合球蛋白（SHBG）基因在人滋养细胞凋亡中的作用。方法用不同siRNA干扰沉默人滋养细胞 SHBG 基因的表达并分为6组，即正常对照组未经转染（Ⅰ组）、空白对照组仅转染脂质体（Ⅱ组）、阴性对照组转染 Nonspecific siRNA（Ⅲ组）、转染组分别转染 SHBGsiRNA-Ⅰ、SHBGsiRNA-Ⅱ、SHBGsiRNA-Ⅲ（Ⅳ、Ⅴ、Ⅵ组），然后通过 Hoechst 33258染色检测人滋养细胞凋亡情况，Real-time PCR 和 Western blot 测定 SHBG mRNA 及蛋白和凋亡相关抗体 Caspase-3 mRNA 及蛋白的表达水平。结果Ⅰ组、Ⅱ组、Ⅲ组 SHBG 和 Caspase-3 mRNA 及蛋白表达水平组间比较差异无统计学意义（P ＞0．05）。Ⅳ组、Ⅴ组、Ⅵ组 SHBG和 Caspase-3 mRNA 及蛋白表达水平组间比较差异无统计学意义（P ＞0．05）。与Ⅰ组、Ⅱ组、Ⅲ组比较，Ⅳ组、Ⅴ组、Ⅵ组 SHBG mR-NA 和蛋白表达量明显下调，Caspase-3 mRNA 及蛋白表达水平升高，滋养层细胞凋亡增加，差异均具有统计学意义（P ＜0．05）。结论采用 siRNA 干扰技术能够降低人滋养细胞中 SHBG 基因表达量，导致人滋养细胞凋亡。%Objective To investigate the effects of sex hormone-binding globulin (SHBG)gene in the apoptosis of human trophoblastic cells.Methods The siRNA specific-targeting SHBG gene was transfected into human trophoblastic cells and they were divided into six groups:trophoblasts without transfection in normal control groups(group Ⅰ);transfect liposome in blank control groups(group Ⅱ);transfect nonspecific siRNA in negative control groups(group Ⅲ);transfect SHBG siRNA-Ⅰ,SHBG siRNA-Ⅱ,SHBG siRNA-Ⅲ respectively in trans-fection group(group Ⅳ,Ⅴ,Ⅵ).Hoechst 33258 dying method was used to detect cell apoptosis.SHBG and Caspase-3 mRNA profiling and the level of SHBG and caspase-3 protein were detected by real-time PCR and Western blot.Results There was no statistical significant difference in the gene expression and protein level of SHBG and caspase-3 in group Ⅰ,Ⅱ and Ⅲ (P >0.05).In Ⅳ,Ⅴ and Ⅵ group,there was no statistical significant difference in the expression level of SHBG and caspase 3 (P >0.05).Compared with group Ⅰ,Ⅱ and Ⅲ,the a-mount of SHBG gene expression decreased obviously,the caspase-3 mRNA and protein level increased obviously and the trophoblast cell ap-optosis increased markedly (P <0.05).Conclusion Through siRNA interference technology can reduce SHBG gene expression in human trophoblastic cells,and it can lead to excessive apoptosis of human trophoblasts cells.