微小核糖核酸-371a-5p靶向调控X相关凋亡抑制蛋白在滋养层细胞凋亡和复发性流产中的作用分析

The role of miR-371a-5p targeted XIAP in apoptosis and recurrent abortion of trophoblastic cells

目的探讨微小核糖核酸-371a-5p(miR-371a-5p)靶向调控X相关凋亡抑制蛋白(XIAP)在滋养层细胞凋亡、复发性流产中的作用。方法滋养层JEG-3细胞经过细胞培养、转染后,分为空白组(Control)、阴性对照组(ND)、miR-371a-5p mimics组及miR-371a-5p inhibitor组。实时荧光定量PCR检测miR-371a-5的表达观察滋养层细胞凋亡能力,免疫印迹(Western blot)检测XIAP、含半胱氨酸的天冬氨酸蛋白水解酶(caspase-3)表达,采用实时荧定量PCR检测XIAP、caspase-3基因。结果Control组miR-371a-5p mRNA表达水平为(1.50±0.09),NC组miR-371a-5p mRNA表达水平为(1.44±0.05),miR-371a-5p mimics组miR-371a-5p mRNA表达水平为(1.23±0.08),miR-371a-5p inhibitor组miR-371a-5p mRNA表达水平为(1.86±0.05),miR-371a-5p mimics组表达低于Control、NC、miR-371a-5pinhibitor组;miR-371a-5p inhibitor组表达高于Control、NC、miR-371a-5p mimics组(F=31.759,P<0.05)。Control组转染48 h后细胞凋亡率为(1.74±0.04)%,NC组转染48 h后细胞凋亡率为(1.76±0.06)%,miR-371a-5p mimics组转染48 h后细胞凋亡率为(3.51±0.09)%,miR-371a-5p inhibitor组转染48 h后细胞凋亡率为(1.33±0.16)%,滋养层细胞转染48 h后miR-371a-5p mimics组细胞凋亡率高于NC、Control及miR-371a-5p inhibitor组;miR-371a-5p inhibitor组低于NC、Control及miR-371a-5p mimics组(F=47.027,P<0.05)。miR-371a-5p mimics组XIAP相对表达量低于NC、Control及miR-371a-5p inhibitor组;miR-371a-5p inhibitor组XIAP相对表达量高于miR-371a-5p mimics、NC及Control组(F=33.541,P<0.05)。miR-371a-5p mimics组caspase-3相对表达量低于NC、Control及miR-371a-5p inhibitor组;miR-371a-5p inhibitor组caspase-3相对表达量高于miR-371a-5p mimics、NC及Control组(F=43.728,P<0.05)。结论miR-371a-5p靶向调控XIAP参与滋养层细胞凋亡的过程,在复发性流产起到重要作用。

Objective The role of micro-ribonucleic acid-371a-5p(miR-371a-5p)targeting X-related apoptosis inhibitory pro-tein(XIAP)in osteosarcoma was studied.Methods Trophoblastic cultured and transfected JEG-3 cells are divided into blank group(Con-trol),negative control group(ND),miR-371 a-5 p micics group,and miR-371a-5p inhibitor group.The expression of real-time fluores-cence quantitative PCR observed trophoblastic apoptosis,immunoimprint(Western blot)detected the expression of aspartate protein hydro-lase containing cysteine(caspase-3),and used real-time screen quantitative PCR to detect the XIAP,caspase-3 gene.Results The ex-pression of miR-371a-5p mRNA in Control group was(1.50±0.09),the expression of miR-371a-5p mRNA in NC group was(1.44±0.05),the expression of miR-371a-5p mRNA in miR-371a-5p mimics group was(1.23±0.08),the expression of miR-371a-5p mRNA in miR-371a-5p inhibitor group was(1.86±0.05),miR-371a-5p mimics group expression is lower than Control,NC,miR-371a-5p in-hibitor group;miR-371a-5p inhibitor group expression is higher than Control,NC,miR-371a-5p mimics group(F=31.759,P<0.05).The apphotic rate after 48 hours in Control group was(1.74±0.04)%,the apphotic rate in NC group was(1.76±0.06)%,the apphotic rate in miR-371a-5p mimics group was(3.51±0.09)%,the apphotic rate inmiR-371a-5p inhibitor group was(1.33±0.16)%,The ap-photic rate of miR-371a-5p mimics after 48 hours was higher than NC,Control,miR-371a-5p inhibitor;miR-371a-5p inhibitor was low-er than NC,Control,miR-371a-5p mimics(F=47.027,P<0.05).The relative expression of the miR-371a-5p mimics group XIAP,caspase-3 is lower than the NC,Control,miR-371a-5p inhibitor group;the miR-371a-5p inhibitor group XIAP,caspase-3 is higher than the miR-371a-5p mimics,NC,Control group(F=33.541,P<0.05).The relative expression of caspase-3 in miR-371a-5p mimics group is lower than that in NC,Control and miR-371a-5p inhibitor group.The relative expression of caspase-3 in miR-371a-5p inhibitor group is higher than that in miR-371a-5p mimics,NC and control groups(F=43.728,P<0.05).Conclusion miR-371a-5p targeted regulation of XIAP is involved in trophoblastic apoptosis and plays an important role in recurrent abortion.