肿瘤坏死因子诱导蛋白8对滋养细胞凋亡和侵袭影响研究  

Effect of TNFAIP8 on apoptosis and invasion of trophoblast cells

作　　者：林容[1] 林靓[1] 李蕊[1] 杨茵[1] 柳之彦 LIN Rong;LIN Liang;LI Rui;YANG Yin;LIU Zhi-yan(Provincial Clinical College of Fujian Medical University,Fujian Provincial Hospital South Branch,Fuzhou 350001,China)

机构地区：[1]福建医科大学省立临床学院福建省立金山医院(福建省立医院南院),福建福州350001

出　　处：《中国实用妇科与产科杂志》2020年第3期267-271,共5页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：福建省自然科学基金计划(2017J01173);福建省卫生厅中青年骨干人才培养项目(2018-ZQN-22);国家自然科学基金青年基金(81800363)。

摘　　要：目的探讨肿瘤坏死因子诱导蛋白8(TNFAIP8)对HTR8/SVneo滋养细胞凋亡和侵袭、迁移的影响。方法应用免疫组化检测2016年11月至2017年6月福建省立医院南院门诊行人工流产术的7~10周单胎正常妊娠妇女30例的正常早孕期绒毛中TNFAIP8的表达情况。构建针对TNFAIP8的慢病毒载体,感染HTR8/SVneo细胞后沉默TNFAIP8的表达,运用qRT-PCR、Western blot检验沉默效果。Transwell实验、细胞划痕实验、FCM法检测沉默TNFAIP8表达后的HTR8/SVneo细胞的侵袭、迁移能力及凋亡的影响。结果 TNFAIP8在正常早孕期绒毛外滋养细胞中阳性表达,且在绒毛组织与外周血淋巴细胞中表达呈正相关。运用qRT-PCR和Western blot检测显示,shTNFAIP8转入HTR8/SVneo细胞后,TNFAIP8蛋白和mRNA水平明显下调(P <0.05)。Transwell实验、细胞划痕实验结果显示沉默HTR8/SVneo细胞中TNFAIP8的表达明显抑制细胞的侵袭和迁移能力(P <0.001)。FCM法检测结果显示,TNFAIP8沉默组细胞凋亡显著增加(P<0.05)。结论在早孕期绒毛外滋养细胞中,阳性表达的TNFAIP8可促进HTR8/SVneo滋养细胞的侵袭及迁移并抑制细胞的凋亡。Objective To explore the effects of TNFAIP8 on the apoptosis,invasion and migration of HTR8/SVneo trophoblast cells.Methods Immunohistochemistry was used to detect the expression of TNFAIP8 in chorionic villi during early pregnancy at 7 to 10 weeks in 30 normal pregnant women with single fetus who underwent artificial abortion in the outpatient department of Fujian Provincial Hospital South Branch between November 2016 and June 2017.Lentiviral vector including TNFAIP8-shRNA was constructed,then it was transfected into HTR8/SV neo cells by virus infection and TNFAIP 8 expression was silenced.The expression levels of TNFAIP8 mRNA and protein in HTR8/SVneo were detected by real-time fluorescent quantitative PCR and Western-blot,respectively.The invasion,migration and apoptosis ability of HTR8/SVneo cells were measured by Transwell,cell scratch assays and FCM after silencing TNFAIP8 expression.Results TNFAIP8 in the cytoplasm of extra villous cytotrophoblast cell early pregnancy represents as positive expression and positive correlation existerd between villous tissue and peripheral blood lymphocytes.2.The results of real-time fluorescent quantitative PCR and Western-blot showed that the expression levels of TNFAIP8 mRNA and protein in HTR8/SVneo cells,which had been infected by TNFAIP8-shRNA,were down-regulated(both P<0.05).The Transwell and cell scratch assays showed that the invasion and migration abilities of HTR8/SVneo cells with TNFAIP8 gene silencing were significantly inhibited(both P<0.001).FCM showed the apoptosis of TNFAIP8 silencing group was significantly increased(P<0.05).Conclusion The positive expression of TNFAIP8 in normal early pregnancy extravillous cytotrophoblast cell promotes the invasion and migration of HTR8/SVneo trophoblast cells and inhibits cell apoptosis.

关 键 词：肿瘤坏死因子诱导蛋白8 滋养细胞 侵袭 迁移 凋亡 

分 类 号：R714.2[医药卫生—妇产科学]