MicroRNA-96对人视网膜色素上皮细胞增殖与迁移的影响

doi:10.3760/cma.j.cn115909-20190327-00095

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Abstract Objective: To investigate the effects of microRNA-96 (miR-96) on the proliferation and migration of human retinal pigment epithelial (RPE) cells. Methods: In this experimental research, the expression of miR-96 was detected in the RPE layer in paraffin sections of the human embryonic eye (20 weeks) by RNA in situ hybridization. MiR-96 or a negative control (NC) was transfected into RPE cells by a lipofectamineRNAiMAX reagent. MTS, flow cytometry, and transwell assays were used to evaluate the effects of miR-96 on cell proliferation, cell cycle, and cell migration. The target gene of miR-96 was predicted by bioinformatics and verified by Western blot. The expression of cell cycle-related proteins in RPE cells was detected by Western blot analysis. Data were analyzed by an independent t test. Results: MiR-96 was highly expressed in human RPE cells. MTS assay showed that the proliferation of RPE cells was inhibited by miR-96 (t=42.174, P=0.002). Flow cytometry revealed that miR-96 resulted in a G1 phase arrest in RPE cells (t=-18.444, P=0.003). Transwell assay also revealed that miR-96 inhibited the migration of RPE cells (t=6.754, P=0.002). Furthermore, MITF was identified as the target gene of miR-96. In addition, the expression of cell cycle-related proteins p-Cdc2, CyclinD2, p-Rb, as well as p-Akt and p-ERK proteins, had significantly decreased in RPE cells transfected with miR-96 (p-Rb: t=11.211, P=0.002; p-Cdc2: t=9.133, P=0.003; CyclinD2: t=7.542, P=0.005; p-ERK: t=16.699, P<0.001; p-Akt: t=23.552, P<0.001). Conclusions: MiR-96 inhibits human RPE cell proliferation and migration by targeting MITF and regulating the expression of cell cycle and migration-related proteins.

Key words： retinal pigment epithelium microRNA-96 cell proliferation cell migration MITF

Received: 17 May 2019

Fund:Zhejiang Provincial Natural Science Foundation of China Grant (LQ17H120009, LQ15H160014)

Corresponding Authors: Dongsheng Yan, State Key Laboratory of Ophthalmology, Optometry and Vision Science, Eye Hospital, Wenzhou Medical University, Wenzhou 325027, China (Email: dnaprotein@hotmail.com)

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