RNA干扰技术抑制SHBG基因表达对滋养细胞凋亡的影响  

Effect of siRNA-mediated SHBG Gene Silencing on Apoptosis of Trophoblasts

作　　者：孙倩[1] 孙磊[1] 金镇[1] 范杰慧[1] 息小雪[1] 

机构地区：[1]中国医科大学附属盛京医院妇产科,沈阳110004

出　　处：《中国医科大学学报》2013年第8期689-692,共4页Journal of China Medical University

基　　金：国家自然科学基金(81170591);沈阳市科学技术计划(F11-262-9-46)

摘　　要：目的探讨RNA干扰技术抑制SHBG基因表达对滋养细胞凋亡的影响。方法在体外通过RNA干扰技术,将SHBGsiRNA转染至原代培养的人绒毛滋养层细胞中,双染流式细胞术检测绒毛滋养层细胞凋亡情况。结果成功原代培养人绒毛滋养层细胞。滋养细胞转染时分成6组:Ⅰ组为正常对照组(未经转染的细胞);Ⅱ组为空白对照组(只转染脂质体,无特异性siRNA);Ⅲ组为阴性对照组(转染Nonspecific siRNA);Ⅳ、Ⅴ、Ⅵ组为转染组(分别转染SHBGsiRNA-Ⅰ、SHBGsiRNA-Ⅱ、SHBGsiRNA-Ⅲ)。成功将SHBGsiRNA转染人绒毛滋养层细胞,结果显示与正常对照Ⅰ组、空白对照Ⅱ组和阴性对照Ⅲ组相比,各转染Ⅳ、Ⅴ、Ⅵ组的滋养层细胞凋亡率明显增加(分别为14.61±3.05、14.37±2.47、16.91±2.61、8.89±0.36、8.64±0.87和9.24±0.60,P<0.05)差异均有统计学意义。结论 siRNA可以有效干扰滋养层细胞SHBG基因的表达,可导致滋养层细胞过度凋亡。Objective To investigate the effect of SHBG gene silencing by RNA interference on the apoptosis of trophoblasts.Methods Small interfering RNA（siRNA）specific-targeting SHBG gene was transfected into cultured human first trimester trophoblasts.Cell apoptosis was detected by the terminal flow cytometer method.Results Trophoblasts were successfully cultured and purified.Trophoblasts were divided into six groups before transfection,group Ⅰ ：normal control groups（trophoblasts without transfection）;group Ⅱ ：blank control groups（transfect liposome,without siRNA）;group Ⅲ：negative control groups（transfect nonspecific siRNA）;group Ⅳ,Ⅴ,Ⅵ：Transfection group（transfect SHBG siRNA-Ⅰ,SHBG siRNA-Ⅱ,SHBG siRNA-Ⅲ respectively）.The SHBG siRNA is successfully transfected into trophoblasts,the apoptotic cell ratio of group IV,group V,group VI were（14.61±3.05,14.37±2.47,16.91±2.61）significantly higher than controls groupⅠ,groupⅡ,groupⅢ（8.89±0.36,8.64±0.87,9.24±0.60）（P 0.05）.Conclusion siRNA can interfere the expression of SHBG gene in trophoblasts effectively,which lead to excessive apoptosis of trophoblasts.

关 键 词：性激素结合球蛋白 滋养层细胞 RNA干扰 细胞凋亡 

分 类 号：R714.2[医药卫生—妇产科学]