miR-152-3p靶向IGF1基因对高糖诱导的ARPE-19细胞活性和凋亡的影响

Effect of miR-152-3p-targeted IGF1 gene on high glucose-induced activity and apoptosis of ARPE-19 cells

目的:探究microRNA-152-3p(miR-152-3p)靶向胰岛素样生长因子1(IGF1)基因对高糖诱导的视网膜色素上皮ARPE-19细胞活性和凋亡的影响,并探讨其作用机制。方法:高糖诱导ARPE-19细胞并转染miR-152-3p mimics,噻唑蓝(MTT)法检测细胞增殖活性,流式细胞术检测细胞凋亡情况,荧光定量PCR(RT-PCR)检测细胞中miR-152-3p水平,蛋白印迹(Western blot)法检测细胞中IGF1和VEGF表达水平,双荧光素酶报告基因检测IGF1和miR-152-3p靶向结合关系。结果:高糖能够降低ARPE-19细胞活性,提高细胞凋亡率,抑制细胞中miR-152-3p的表达,提高IGF1和VEGF的表达;而过表达miR-152-3p能够回调高糖诱导的细胞活性抑制及凋亡增加,抑制IGF1和VEGF的表达。双荧光素酶报告基因实验验证了IGF1是miR-152-3p的靶基因。结论:miR-152-3p可通过靶向IGF1基因调节VEGF的表达抑制高糖诱导的ARPE-19细胞活性抑制和凋亡增加。

AIM:To investigate the effect of microRNA-152-3p (miR-152-3p) targeting insulin-like growth factor 1 (IGF1) gene on high glucose-induced retinal pigment epithelial ARPE-19 cell activity and apoptosis, and to explore its role mechanism. METHODS: High glucose was induced into ARPE-19 cells and transfected with miR-152-3p mimics. MTT assay was used to detect cell proliferation activity. Flow cytometry was used to detect apoptosis. Fluorescence quantitative PCR (RT-PCR) was used to detect cells. The expression levels of IGF1 and VEGF in the cells were detected by Western blot and the binding relationship between IGF1 and miR-152-3p was detected by the dual luciferase reporter gene. RESULTS:High glucose can decrease the activity of ARPE-19 cells, increase the apoptosis rate, inhibit the expression of miR-152-3p and increase the expression of IGF1 and VEGF. Over expression of miR-152-3p can up-regulate high glucose-induced cells. Increased activity and increased apoptosis inhibited the expression of IGF1 and VEGF. The dual luciferase reporter gene assay verified that IGF1 is the target gene of miR-152-3p. CONCLUSION: miR-152-3p can inhibit the inhibition of high glucose-induced ARPE-19 cell activity and increase apoptosis by targeting IGF1 gene.