lncRNA MALAT1调节miR-155-5p/IGF2轴对高糖诱导的滋养层细胞的增殖、迁移和侵袭的影响

Influences of lncRNA MALAT1 on the proliferation, migration and invasion of high glucose-induced trophoblast cells by regulating the miR-155-5p/IGF2 axis

目的 探讨长链非编码RNA肺腺癌转移相关转录本1(lncRNAMALAT1)通过调节微小RNA-155-5p(miR-155-5p)/胰岛素样生长因子2(IGF2)轴对滋养层细胞增殖、迁移和侵袭的影响。方法 体外培养人绒毛膜滋养层细胞HTR8/SVneo,待其分化成熟后分为6组:正常组、高糖组、si-NC组、si-MALAT1组、si-MALAT1+anti-miR-NC组、si-MALAT1+anti-miR-155-5p组。qRT-PCR实验检测lncRNA MALAT1、miR-155-5p、IGF2 mRNA表达水平;MTT、Transwell和划痕愈合实验检测细胞增殖、迁移和侵袭情况;Western blot法检测IGF2、MMP-2、MMP-9蛋白表达。通过双荧光素酶报告基因检测验证lncRNAMALAT1、IGF2和miR-155-5p的靶向关系。结果 与正常组比较,高糖组lncRNA MALAT1、IGF2 mRNA和蛋白表达明显升高,miR-155-5p表达明显下调,细胞增殖、迁移和侵袭能力及MMP-2、MMP-9蛋白表达降低(P<0.05);沉默lncRNA MALAT1能够显著提高HTR8/SVneo细胞增殖、迁移和侵袭能力,上调miR-155-5p表达,下调IGF2 mRNA和蛋白表达,升高MMP-2、MMP-9蛋白表达(P<0.05);在沉默lncRNA MALAT1的基础上抑制miR-155-5p表达能够部分逆转上述指标水平(P<0.05);双荧光素酶报告基因检测结果证实lncRNA MALAT1可以靶向调节miR-155-5p/IGF2轴。结论 沉默lncRNAMALAT1能够通过靶向调节miR-155-5p/IGF2轴,促进高糖诱导的滋养层细胞增殖、迁移和侵袭。

Objective To investigate the influences of long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1(lncRNA MALAT1) on proliferation, migration and invasion of trophoblast cells by regulating the microRNA-155-5p(miR-155-5p)/insulin-like growth factor 2(IGF2) axis. Methods Human chorionic trophoblast cells HTR8/SVneo were cultured in vitro and divided into six groups after differentiation and maturity: Normal group, high glucose group, si-NC group, si-MALAT1 group, si-MALAT1+anti-miR-NC group, and si-MALAT1+anti-miR-155-5p group. qRT-PCR experiment was used to measure the expression levels of lncRNA MALAT1, miR-155-5p and IGF2 mRNA. MTT, Transwell and scratch healing assays were performed to detect the cell proliferation, migration and invasion;the expression of IGF2,MMP-2 and MMP-9 protein were detected by Western blot, the targeting relationship of lncRNA MALAT1, IGF2 and miR-155-5p were verified by dual-luciferase reporter gene assay. Results Compared with the normal group, the expression of lncRNA MALAT1 and the mRNA and protein expression of IGF2 in the high glucose group was significantly increased, the expression of miR-155-5p was significantly down-regulated, the abilities of cell proliferation, migration and invasion, and the expression of MMP-2 and MMP-9 proteins decreased(P<0.05). Silencing lncRNA MALAT1 was able to significantly improve the proliferation, migration and invasion abilities of HTR8/SVneo cells, up-regulate the expression of miR-155-5p,down-regulate the mRNA and protein expression of IGF2, and increase the protein expression of MMP-2 and MMP-9(P<0.05).Inhibiting the expression of miR-155-5p on the basis of silencing lncRNA MALAT1 can partially reverse the levels of the above indicators(P<0.05). The results of dual-luciferase reporter gene assay confirmed that the lncRNA MALAT1 was able to target and regulate the miR-155-5p/IGF2 axis. Conclusion Silencing lncRNA MALAT1 can promote high glucose-induced proliferation, migration and invasion of trophoblast cells by targeting the regulation of miR-155-5p/IGF2 axis.