CircACTR2调节miR-497-5p/TXNIP轴对高糖诱导的滋养层细胞增殖、凋亡和上皮间质转化的影响

Effect of CircACTR2 on high glucose induced proliferation,apoptosis,and epithelial mesenchymal transition of trophoblast cells by regulating the miR-497-5p/TXNIP axis

目的探讨CircACTR2调节miR-497-5p/TXNIP轴对高糖诱导的滋养层细胞增殖、凋亡和上皮间质转化(EMT)的影响。方法将HTR-8/SVneo细胞分为control组和高糖组,并对高糖组细胞进行质粒转染。qRT-PCR检测胎盘组织和HTR-8/SVneo细胞中CircACTR2、miR-497-5p、TXNIP mRNA表达,CCK-8法测定细胞增殖,流式细胞术检测细胞凋亡,Transwell小室检测细胞迁移和侵袭,Western blot检测TXNIP、Bax、Bcl-2、E-cadherin、N-cadherin蛋白表达,双荧光素酶报告实验与RIP实验验证CircACTR2、TXNIP与miR-497-5p的靶向调控关系。结果CircACTR2、TXNIP在GDM胎盘组织和高糖诱导的HTR-8/SVneo细胞中表达升高,miR-497-5p表达降低(P<0.05);与control组比较,高糖组HTR-8/SVneo细胞OD_(450)值、迁移、侵袭细胞数、Bcl-2、N-cadherin表达显著降低,凋亡率、Bax、E-cadherin表达显著升高(P<0.05);干扰CircACTR2或过表达miR-497-5p可显著促进HTR-8/SVneo细胞增殖、迁移、侵袭和EMT,抑制凋亡(P<0.05);抑制miR-497-5p表达或过表达TXNIP可逆转干扰CircACTR2或过表达miR-497-5p对HTR-8/SVneo细胞增殖、迁移、侵袭和EMT的促进作用,增加凋亡(P<0.05)。双荧光素酶报告与RIP实验显示CircACTR2、TXNIP与miR-497-5p存在靶向调控关系。结论CircACTR2在高糖诱导的HTR-8/SVneo细胞中表达上调,干扰CircACTR2表达可能通过调节miR-497-5p/TXNIP,促进细胞增殖、迁移、侵袭和EMT,抑制凋亡。

Objective To investigate the impacts of CircACTR2 on high glucose induced proliferation,apoptosis,and epithelial mesenchymal transition(EMT)of trophoblast cells by regulating the miR-497-5p/TXNIP axis.Methods HTR-8/SVneo cells were grouped into control group and high glucose group,and cells in the high glucose group were transfected with plasmids.qRT-PCR was applied to detect the expression of CircACTR2,miR-497-5p,and TXNIP mRNA in placental tissue and HTR-8/SVneo cells,CCK-8 method was applied to measure cell proliferation,flow cytometry was applied to detect cell apoptosis,Transwell cells were used to detect cell migration and invasion,Western blot was applied to detect the expression of TXNIP,Bax,Bcl-2,E-cadherin,N-cadherin proteins,in addition,dual luciferase reporter and RIP experiments were applied to verify the targeted regulatory relationship between CircACTR2,TXNIP,and miR-497-5p.Results The expression of CircACTR2 and TXNIP increased in GDM placental tissue and high glucose induced HTR-8/SVneo cells,while the expression of miR-497-5p decreased(P<0.05);compared with the control group,the OD_(450) value of HTR-8/SVneo cells,migration and invasion cell counts,expression of Bcl-2,N-cadherin in the high glucose group were obviously reduced,the apoptosis rate,the expression of Bax and E-cadherin were obviously increased(P<0.05);interference with CircACTR2 or overexpression of miR-497-5p was able to obviously promote the proliferation,migration,invasion,and EMT of HTR-8/SVneo cells,and inhibit apoptosis(P<0.05);inhibiting the expression of miR-497-5p or overexpressing TXNIP was able to reverse the inhibitory effects of CircACTR2 or overexpressing miR-497-5p on the proliferation,migration,invasion,and EMT of HTR-8/SVneo cells,and increase apoptosis(P<0.05).The dual luciferase reporter and RIP experiments showed that CircACTR2,TXNIP and miR-497-5p had a targeted regulatory relationship.Conclusion CircACTR2 is up-regulated in high glucose induced HTR-8/SVneo cells,and interference with CircACTR2 expression may promote cell proliferation,migration,invasion,EMT,and inhibit apoptosis by regulating miR-497-5p/TXNIP.