ARID1A通过阻断Wnt/β-catenin通路抑制滋养层细胞增殖、迁移和侵袭

ARID1A inhibits trophoblast cell proliferation,migration and invasion via blocking Wnt/β-catenin signaling pathway

目的探讨富含AT的相互作用结构域蛋白1A(AT-rich interactive domain 1A,ARID1A)对子痫前期(preeclampsia,PE)滋养层细胞增殖、侵袭及迁移能力的影响及潜在分子机制。方法采用RT-qPCR及Western blot测定30例PE患者及30例正常妊娠者胎盘组织中ARID1A表达情况。以人绒毛滋养层细胞HTR-8/SVneo为研究对象,实验分组为:control组(无处理)、NC组(转染空载体)、ARID1A组(转染ARID1A过表达载体)、ARID1A+氯化锂(LiCl)组(转染ARID1A过表达载体后再用50 mmol/L的LiCl处理)。采用MTT法检测细胞增殖,采用Transwell实验检测细胞侵袭能力和迁移能力,采用RT-qPCR及Western blot法检测胎盘组织中ARID1A的mRNA和蛋白表达水平,采用Western blot法检测HTR-8/SVneo细胞中ARID1A、MMP-2、MMP-9、Wnt1、β-catenin和Cyclin D1的蛋白表达水平。结果PE患者胎盘组织中ARID1A的表达水平显著高于正常胎盘组织(P<0.05)。与control组相比,ARID1A组细胞增殖、侵袭、迁移能力显著降低(P<0.05),MMP-2、MMP-9、Wnt1、β-catenin和Cyclin D1蛋白表达水平显著降低(P<0.05)。与ARID1A组相比,ARID1A+LiCl组Wnt1、β-catenin和Cyclin D1蛋白表达水平显著升高(P<0.05),细胞侵袭和迁移能力显著升高(P<0.05)。结论ARID1A在PE胎盘组织中高表达,ARID1A可能通过Wnt/β-catenin信号通路抑制滋养层细胞的侵袭和迁移。

Objective To elucidate the effect and its potential molecular mechanism of AT-rich interactive domain 1A(ARID1A)on cell proliferation,migration and invasion in human trophoblast cells.Methods RT-qPCR and Western blot were conducted to examine the expression level of ARID1A in placenta tissues from 30 preeclampsia(PE)and 30 normal pregnant women.Human trophoblast cell line HTR-8/SVneo was used in this study.Cells were divided into control group(no any treatment),NC group(transfected with empty vector),ARID1A group(transfected with ARID1A overexpression vector)and ARID1A+LiCl group(transfected with ARID1A overexpression vector and then treated with 50 mmol/L LiCl).MTT assay was carried out to evaluate cell proliferation.Transwell assay was applied to estimate the cell invasion and migration.RT-qPCR and Western blot assay were used to detect the mRNA and protein levels of ARID1A in placenta tissues,respectively.Western blot assay was used to measure the protein levels of ARID1A,MMP-2,MMP-9,Wnt1,β-catenin and Cyclin D1 in HTR-8/SVneo cells.Results The expression level of ARID1A in PE tissues was significantly higher than that in normal placental tissues(P<0.05).Compared with control group,the cell proliferation,invasion and migration abilities were significantly downregulated in ARID1A group(P<0.05).Compared with control group,the protein expression levels of MMP-2,MMP-9,Wnt1,β-catenin and Cyclin D1 were also significantly downregulated in ARID1A group(P<0.05).Compared with ARID1A group,the protein expression levels of Wnt1,β-catenin and Cyclin D1 were significantly upregulated in ARID1A+LiCl group(P<0.05).Compared with ARID1A group,the cell invasion and migration abilities were also significantly upregulated in ARID1A+LiCl group(P<0.05).Conclusion ARID1A is highly expressed in placental tissues of PE.ARID1A might inhibit the cell invasion and migration of trophoblast cells via Wnt/β-catenin signaling pathway.