干扰小RNA沉默CAV1表达对人绒毛膜癌JEG-3细胞侵袭、迁移能力的影响及其作用机制研究

目的 探究干扰小RNA(siRNA)沉默小窝蛋白-1(CAV1)基因表达对人绒毛膜癌JEG-3细胞侵袭、迁移能力的影响及其可能的作用机制.方法 将人绒毛膜癌JEG-3细胞分为对照组(不进行转染)、阴性组(转染siRNA-NC)和siRNA-CAV1组(转染siRNA-CAV1).Transwell法检测下调CAV1表达对细胞侵袭、迁移能力的影响;实时定量PCR(qRT-PCR)检测转染细胞中CAV1 mRNA的表达水平;蛋白质印迹法(Western blot)检测细胞中CAV1、丝苏氨酸蛋白激酶(AKT)、雷帕霉素靶蛋白(MTOR)、核糖体p70S6激酶(p70S6K)、磷酸化AKT(p-AKT)、磷酸化MTOR(p-MTOR)、磷酸化p70S6K(p-p70S6K)蛋白的表达水平.结果 siRNA-CAV1组JEG-3细胞中CAV1 mRNA和蛋白的相对表达量均低于对照组(P﹤0.05);siRNA-CAV1组JEG-3细胞的侵袭数目、迁移数目均低于对照组(P﹤0.05);siRNA-CAV1组JEG-3细胞中AKT、MTOR、p70S6K以及磷酸化水平均低于对照组(P﹤0.05).结论 沉默CAV1表达可以抑制人绒毛膜癌JEG-3细胞的侵袭和迁移能力,该作用与AKT/MTOR/p70S6K信号通路有关.

Effect and mechanism of small interfering RNA silencing CAV1 expression on the invasion and migration of human choriocarcinoma JEG-3 cells

Objective To investigate the effect and mechanism of small interfering RNA (siRNA) silencing caveolin-1 (CAV1) gene expression on the invasion and migration of human choriocarcinoma JEG-3 cells. Method The human cho-riocarcinoma JEG-3 cells were divided into three groups:control group (without transfected), negative group (transfected by siRNA-NC) and siRNA-CAV1 group (transfected by siRNA-CAV1). Cell migration and invasion effected by down-regulated expression of CAV1 were detected by Transwell assay;the level of CAV1 mRNA in transfected cells was detect-ed by qRT-PCR;the expression of CAV1, serine/threonine kinase (AKT), mechanistic target of rapamycin (MTOR), p70 ribosomal protein S6 kinase (p70S6K), phosphorylated AKT (p-AKT), phosphorylated MTOR (p-MTOR), phosphorylat-ed p70S6K (p-p70S6K) were detected by Western blot. Result After transfected, the expression of CAV1 protein and mRNA in siRNA-CAV1 group was lower than that in the control group (P<0.05);the amount of invasive cells and migrat-ed cells of JEG-3 cells in siRNA-CAV1 group were lower than that in the control group (P<0.05);compared to the con-trol group, protein expression and phosphorylation levels of AKT, MTOR and p70S6K all decreased in siRNA-CAV1 group (P<0.05). Conclusion Silenced expression of CAV1 could inhibit the invasion and migration ability of choriocarci-noma JEG-3 cell line, which may related to the AKT/MTOR/p70S6K signaling pathway.