lncRNA XIST靶向miR-182-5p/HIF-2α分子轴对滋养层细胞生物学行为特性影响的机制研究OA

Objective To investigate the molecular mechanism by which long-chain non-coding RNA(lncRNA)X-inactive specific transcript(XIST)regulates the biological activity of trophoblast cells.Methods Human HTR-8/Svneo trophoblast cells were cultured in vitro and separated into control,interference empty(transfected with small interfering RNA-normal control(si-NC)),si-XIST-1(transfected with si-XIST-1),si-XIST-1+anti-NC(co-transfected with si-XIST-1 and anti-NC),and si-XIST-1+anti-182-5p(co-transfected with si-XIST-1 and anti-182-5p)groups.XIST,miR-182-5p,and hypoxia inducible factor-2α(HIF-2α)mRNA expression levels were measured by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Cell proliferation was detected by Cell Counting Kit-8(CCK-8)and 5-ethynyl-2'-deoxyuridine(EdU)assays.Apoptosis was detected by flow cytometry.Cell migration and invasion abilities were measured by scratch-healing and Transwell experiments.HIF-2α,cleaved-caspase-3,Bcl-2,Bax,matrix metalloproteinase(MMP)-2,and MMP-9 protein levels in cells were measured by Western blot.The targeting relationship of miR-182-5p with XIST and HIF-2α was verified by dual-luciferase reporter assay.Results XIST and HIF-2α mRNA levels,apoptosis rate,and HIF-2α,cleaved-caspase-3,and Bax protein levels in the si-XIST-1 group were lower than those in the control and interference empty groups,while miR-182-5p,cell viability,EdU-positive rate,scratch-healing rate,number of invaded cells,and Bcl-2,MMP-2,and MMP-9 protein levels were higher(P＜0.05).HIF-2α mRNA level,apoptosis rate,and HIF-2α,cleaved-caspase-3,and Bax protein levels in the si-XIST-1+anti-182-5p group were higher than those in the si-XIST-1 and si-XIST-1+anti-NC groups,while miR-182-5p,cell viability,EdU-positive rate,scratch-healing rate,number of invaded cells,and Bcl-2,MMP-2,and MMP-9 protein levels were lower(P＜0.05).LncRNA XIST was able to target miR-182-5p in HTR-8/Svneo cells,and HIF-2α was the target of miR-182-5p.Conclusions Knocking-down lncRNA XIST may inhibit HIF-2α expression by competitively binding to miR-182-5p,thereby enhancing the proliferation,migration,and invasion abilities of HTR-8/Svneo cells and inhibiting cell apoptosis.