lncRNA DANCR加重低氧诱导的滋养细胞损伤

lncRNA DANCR aggravates hypoxia-induced trophoblast injury

目的:探究长链非编码RNA(lncRNA)DANCR在低氧诱导的滋养细胞损伤中的作用及其相关机制。方法:常氧和低氧条件下体外培养人胎盘滋养细胞系(HTR-8/SVneo)。将si-lncRNA DANCR和si-miR-33b转染至HTR-8/SVneo细胞,低氧条件培养,分组:si-NC组、si-lncRNA DANCR组和si-lncRNA DANCR+si-miR-33b组。qRT-PCR法检测细胞lncRNA DANCR和miR-33b表达水平,Western blot检测细胞Beclin-1、p62和γH2AX蛋白表达水平,彗尾实验检测细胞损伤水平。采用荧光素酶报告基因实验检测lncRNA DANCR和miR-33b靶向性。结果:与常氧组相比,低氧组细胞中lncRNA DANCR表达水平升高,Beclin-1和γH2AX蛋白表达升高,p62蛋白表达降低,彗尾长度增加(P<0.05)。与si-NC组相比,si-lncRNA DANCR组细胞中lncRNA DANCR表达水平降低,miR-33b表达升高,Beclin-1和γH2AX蛋白表达降低,p62蛋白表达升高,彗尾长度减少(P<0.05)。HTR-8/SVneo细胞中lncRNA DANCR靶向miR-33b,与si-lncRNA DANCR组相比,si-lncRNA DANCR+si-miR-33b组细胞中miR-33b表达水平降低,Beclin-1和γH2AX蛋白表达升高,p62蛋白表达降低,彗尾长度增加(P<0.05)。结论:lncRNA DANCR通过靶向抑制miR-33b表达,加重低氧诱导的滋养细胞损伤。

Objective:To investigate the role of long non-coding RNA(lncRNA)DANCR in hypoxia-induced trophoblast injury and its associated mechanisms.Methods:Human placental trophoblast cell line(HTR-8/SVneo)was cultured in vitro under normoxic and hypoxic conditions.The HTR-8/SVneo cells were transfected with si-lncRNA DANCR and si-miR-33b,then cultured under hypoxia,and divided into si-NC group,si-lncRNA DANCR group,and si-lncRNA DANCR+si-miR-33b group.The expression levels of lncRNA DANCR and miR-33b were detected by qRT-PCR.The protein expression levels of Beclin-1,p62,andγH2AX were measured by Western blot.The DNA damage level was assessed using the comet assay.The targeting relationship between lncRNA DANCR and miR-33b was examined using a luciferase reporter assay.Results:Compared with the normoxia group,the expression levels of lncRNA DANCR,Beclin-1,andγH2AX were elevated in the hypoxia group cells,whereas the expression of p62 was reduced,and the comet tail length increased(P<0.05).Compared with the si-NC group,the expression level of lncRNA DANCR decreased in the si-lncRNA DANCR group cells,miR-33b expression increased,the expressions of Beclin-1 andγH2AX decreased,p62 expression increased,and the comet tail length shortened(P<0.05).lncRNA DANCR targets miR-33b in HTR-8/SVneo cells.Compared with the si-lncRNA DANCR group,the expression level of miR-33b decreased in the si-lncRNA DANCR+si-miR-33b group cells,the expressions of Beclin-1 andγH2AX increased,p62 expression decreased,and the comet tail length extended(P<0.05).Conclusion:lncRNA DANCR aggravates hypoxia-induced trophoblast injury by targeting the suppression of miR-33b expression.