miR-519a通过调节PTEN促进滋养层细胞增殖迁移侵袭能力影响巨大儿的产生

miR-519a targets on PTEN to promote trophoblast proliferation and invasion to affect fetal macrosomia

目的检测miR-519a在非糖尿病巨大儿胎盘的表达情况,探讨miR-519a对滋养层细胞增殖和侵袭的影响及可能机制。方法收集非糖尿病巨大儿和正常出生体重儿胎盘组织,qRT-PCR方法检测组织中miR-519a的表达情况;体外培养人滋养层细胞系HTR-8/SVneo,并转染miR-519a模拟物和抑制剂,qRT-PCR检测miR-519a在细胞系中的表达情况;CCK-8实验检测HTR-8/SVneo细胞增殖变化;Transwell实验检测细胞侵袭情况;荧光素酶基因报告检测miR-519a与PTEN的3’UTR区结合情况;Western blot检测PTEN在HTR-8/SVneo细胞和巨大儿胎盘组织中的表达。结果与正常体重儿相比,miR-519a在巨大儿胎盘组织中表达水平显著上调。转染miR-519a模拟物可促进HTR8/SVneo细胞增殖和侵袭转移;荧光素酶报告基因实验表明miR-519a与PETN的3’UTR区靶向结合;同时发现在巨大儿胎盘组织中miR-519a和PTEN均存在负相关。结论miR-519a可作为非糖尿病巨大儿潜在的诊断标记物,通过靶向PETN蛋白促进滋养层细胞增殖和侵袭,提示其与巨大儿的产生密切相关。

Objective To observe the expression of miR-519 a in placenta of non-diabetic fetal macrosomia,to explore the effects of miR-519 a on the proliferation and invasion of trophoblast cells.Methods The expression of miR-519 a in placenta was detected in non-diabetic fetal macrosomia and normal birth weight infants by qRT-PCR.The experiment human trophoblast cell line HTR-8/SVneo in vitro was transfected by miR-519 a mimics and inhibitors,and the expression was confirmed by qRT-PCR.CCK8 assay was used to detect cell proliferation;Transwell assay was used to detect cell invasion;Luciferase report assay was used to detect the binding of miR-519 a and PTEN 3’UTR sequence.Western blot were used to detect the expression of PTEN in placenta.Results The expression level of miR-519 a was increased in the placenta of fetal macrosomia.The overexpression of miR-519 a improved the proliferation and invasion of HTR-8/SVneo cells.The luciferase report assay showed that miR-519 a could bind to 3’UTR of PTEN.The expression of PETN and miR-519 a was negative correlation in placenta of fetal macrosomia.Conclusion miR-519 a can be used as a potential diagnostic marker for non-diabetic fetal macrosomia,MiR-519 a promotes trophoblast cell proliferation and invasion by targeting PETN protein,miR-519 a is related to fetal macrosomia.