人胎盘滋养细胞胞内双链DNA识别受体的表达模式及其在介导滋养细胞死亡中的作用

The expression pattern of intracellular double-stranded DNA sensor in human placental trophoblast cells and its role in trophoblast cells death

目的探讨人胎盘滋养细胞胞内双链DNA识别受体的表达模式及其在介导滋养细胞死亡中的作用,揭示滋养细胞在胎盘免疫学屏障功能中的固有免疫识别作用.方法利用RT-PCR方法检测胞内双链DNA识别受体在4株人滋养细胞系HTR-8/SVneo、JEG3、JAR、BeWo中的表达情况;利用不同浓度人工合成双链DNA模拟物poly(dA:dT)刺激人滋养细胞系JEG3,MTT法检测细胞活力;为明确胞内双链DNA诱导的滋养细胞死亡方式,MTT法检测poly(dA:dT)刺激的JEG3细胞应用程序性细胞死亡特异性抑制剂处理24h后的存活情况.结果人滋养细胞系HTR-8/SVneo、JEG3、JAR、BeWo可检测到胞内双链DNA识别受体IFI16、AIM2、DHX9、DHX36、LRRFIP1和KU70的mRNA表达,但未检测到ZBP1/DAI的表达;利用不同浓度poly(dA:dT)转染JEG3细胞系后,JEG3细胞以剂量依赖的方式发生细胞死亡;poly(dA:dT)诱导的细胞死亡可以被加入的细胞凋亡抑制剂z-VAD-fmk、细胞焦亡抑制剂VX-765部分逆转,程序性坏死抑制剂Necrostatin-1则不能影响双链DNA介导的细胞死亡.结论人胎盘滋养细胞表达多种胞内双链DNA识别受体,能够感受双链DNA刺激,以剂量依赖的方式介导滋养细胞发生细胞凋亡、细胞焦亡,但不发生细胞程序性坏死.

Objective To investigate the expression pattern of intracellular double-stranded DNA sensor in human placental trophoblast cells and its role in trophoblast cells death,to reveal the role of trophoblast cells in placental immunological barrier.Methods Analysed the expression of intracellular double-stranded DNA sensors in four human trophoblast cells lines(HTR-8/SVneo、JEG3、JAR、BeWo)by RT-PCR method.Crystal violet stain and MTT analysis were used to evaluate trophoblast cells death after poly(dA:dT)stimulation.Results RT-PCR results showed that four human trophoblast cell lines expressed IFI16,AIM2,DHX9,DHX36,LRRFIP1 and KU70,but no expression of ZBP1/DAI was detected.Double-stranded DNA induced cell death in a dose-dependent manner,and could be partially rescued by z-VAD-fmk and VX-765,but not Necrostain-1.Conclusion Human placental trophoblast cells express multiple intracellular double-stranded DNA sensors,which can recognize double-stranded DNA stimulation and induce apoptosis and pyroptosis in a dose-dependent manner,but no programmed cell necrosis.