膜蛋白CD81增强滋养细胞的黏附能力及机制研究

CD81 enhances the adhesion of trophoblasts and related mechanisms

目的探讨CD81对滋养细胞黏附能力的影响及可能机制,以便阐明CD81参与子痫前期发病的具体机制。方法体外培养永生化的人孕早期绒毛外滋养细胞系HTR-8/SVneo细胞,并感染高表达CD81的腺病毒(Ad-CD81)或转染抑制内源性CD81的干扰RNA(si-CD81)。采用细胞黏附实验检测HTR-8/SVneo细胞的黏附能力,采用免疫印迹法检测HTR-8/SVneo细胞中黏着斑激酶(FAK)的表达情况。结果与感染Ad-CTL的细胞相比,感染Ad-CD81的HTR-8/SVneo细胞黏附能力增强,570 nm处的光密度值显著增加,差异有统计学意义(P<0.05)。与转染si-CTL的细胞相比,转染si-CD81的HTR-8/SVneo细胞黏附能力减弱,570 nm处的光密度值显著下降,差异有统计学意义(P<0.05)。随着Ad-CD81浓度的增加,感染Ad-CD81的HTR-8/SVneo细胞中FAK的蛋白表达较Ad-CTL感染细胞逐渐下降(P<0.05或P<0.01)。随着si-CD81浓度的增加,转染si-CD81的HTR-8/SVneo细胞中FAK的蛋白表达较si-CTL转染细胞逐渐增加(P<0.05或P<0.01)。结论膜蛋白CD81可增强滋养细胞的黏附能力,其分子机制可能与CD81抑制滋养细胞中FAK的蛋白表达有关。

Objective To explore the effect of CD81 on trophoblast adhesion and related mechanisms,in order to determine the role of CD81 in the pathogenesis of preeclampsia.Methods A human first-trimester-derived trophoblast cell line(HTR-8/SVneo cells)was cultivated in vitro before transfection with adenovirus Ad-CD81 which highly ex-pressed CD81 or si-CD81 which inhibited endogenous CD81 through interference RNA.Then,the ability of cell adhesion was detected by adhesion assay.The expression of focal adhesion kinase(FAK)in HTR-8/SVneo cells was detected by Western blot.Results Compared with Ad-CTL-infected cells,Ad-CD81-infected HTR-8/SVneo cells showed en-hanced adhesion ability and increased optical density at 570 nm(P<0.05).Compared with si-CTL-transfected cells,si-CTL-transfected HTR-8/SVneo cells presented weakened adhesion ability and decreased optical density at 570 nm(P<0.05).Furthermore,with the increase of concentration gradients,the expression of FAK was down-regulated in the Ad-CD81-infected HTR-8/SVneo cells,but up-regulated in the si-CD81-transfected cells(P<0.05 or P<0.01).Con-clusions CD81 can enhance the adhesion of trophoblasts,which may be associated with its inhibitory effect on FAK ex-pression in trophoblasts.