四次跨膜蛋白CD81促进人绒毛外滋养细胞失巢凋亡的初步研究  

Tetraspanin CD81 promotes the anoikis of human extravillous trophoblast cells.

作　　者：沈莉[1] 刁振宇[1] 薛平平[2] 龚萍[3] 刘沫[2] 胡娅莉[1] 

机构地区：[1]南京大学医学院附属鼓楼医院妇产科,南京210008 [2]南京医科大学鼓楼临床医学院,南京210029 [3]南京中医药大学,南京210023

出　　处：《现代妇产科进展》2013年第9期700-703,共4页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金(No:81070508)

摘　　要：目的:探讨四次跨膜蛋白CD81表达对人妊娠早期绒毛外滋养细胞失巢凋亡的作用。方法:构建CD81的高表达质粒(pCS2-CD81),将pCS2-CD81或CD81特异的小干扰RNA(siCD81)瞬时转染至人妊娠早期绒毛外滋养细胞(HTR-8/SVneo细胞),同时转染pCS2-EV或siRNA control作为对照组。实时荧光定量PCR(QRT-PCR)和Western blot法检测pCS2-CD81及siCD81在HTR-8/SVneo细胞中的表达效率。采用聚甲基丙烯酸2-羧乙基酯(poly-HEMA)包被的培养皿模拟细胞悬浮培养的环境,采用Annexin V/PI双染法分别检测CD81对正常贴壁生长及悬浮培养的HTR-8/SVneo细胞凋亡的影响。结果:成功构建CD81的高表达质粒(pCS2-CD81)。与转染pCS2-EV比较,瞬时转染pCS2-CD81的HTR-8/SVneo细胞高表达CD81蛋白(P<0.05);与转染siRNA control比较,siCD81有效抑制HTR-8/SVneo细胞的内源性CD81表达(P<0.05)。过表达CD81或抑制内源性CD81表达对HTR-8/SVneo细胞的普通凋亡无显著影响,但过表达CD81可显著促进HTR-8/SVneo细胞的失巢凋亡(P<0.05),抑制内源性CD81表达能减少HTR-8/SVneo细胞的失巢凋亡达15.5%(P<0.05)。结论:四次跨膜蛋白CD81促进人妊娠早期绒毛外滋养细胞的失巢凋亡。Objective： To explore the functional role of tetraspanin CD81 in human extravillous trophoblast cell anoikis.Methods： We constructed the pCS 2-CD81 plasmid.The expression of CD81 was assessed by quantitative real-time PCR（QRT-PCR） and Western blot analysis in HTR-8 / SV neo cells（a human first-trimester-derived trophoblast cell line） transfected with pCS 2-CD81 or CD81-specific siRNA（siCD81）.We examined the effect of CD81 on HTR-8 / SV neo cell apoptosis by flow cytometry（Annexin V / Propidium Iodide double staining） when the cells were cultured under normal condition.To imitate cell suspended culture condition,we cultured HTR-8 / SV neo cells in poly（2-hydroxyethyl methacrylate）（poly-HEMA）-covered plates and examined the effect of CD81 on cell anoikis by flow cytometry.Results： We obtained the pCS 2-CD81 plasmid,which could express higher CD81 protein in pCS 2 CD81-transfected cells than in pCS 2-EV-transfected cells（2.94 fold,P0.05）.CD81 did not influence the apoptosis of HTR-8 / SV neo cells cultured under normal condition.Over-expression of CD81 enhanced HTR-8 / SV neo cell anoikis（P 0.05）,and inhibition of endogenous CD81 significantly reduced HTR-8 / SV neo cell anoikis by 15.5%（P 0.05）.Conclusion： Tetraspanin CD81 promotes the anoikis of human extravillous trophoblast cells.

关 键 词：四次跨膜蛋白CD81 HTR-8 SVneo细胞 失巢凋亡 

分 类 号：R714.2[医药卫生—妇产科学]