miR-30-5p通过下调FOXG1表达抑制视网膜母细胞瘤细胞增殖

doi:10.6040/j.issn.1673-3770.0.2021.466

摘要/Abstract

摘要： 目的探讨miR-30-5p对视网膜母细胞瘤细胞增殖的影响及作用机制。方法采用实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-30-5p在视网膜母细胞瘤细胞系和人视网膜内皮细胞(HRECs)中的表达。利用TargetScan数据库进行生物信息学分析并预测miR-30-5p靶基因,双荧光素酶报告基因实验验证miR-30-5p与FOXG1的3'UTR结合能力及靶向关系,qRT-PCR和Western blotting分别检测miR-30-5p对FOXG1的mRNA与蛋白表达影响。瞬时转染Y79细胞后,通过CCK8法检测各组Y79细胞的增殖。结果与HRECs比较,miR-30-5p在视网膜母细胞瘤细胞中表达降低,FOXG1在视网膜母细胞瘤细胞中表达升高。生物信息学预测结果显示miR-30-5p与FOXG1存在结合位点,qRT-PCR和Western blotting显示miR-30-5p可负调控FOXG1的mRNA和蛋白的表达,双荧光素酶实验结果证实miR-30-5p与FOXG1 3'UTR存在靶向关系。瞬时转染miR-30-5p可升高Y79细胞中miR-30-5p的表达水平,抑制Y79细胞的增殖活力。过表达FOXG1可逆转miR-30-5p上调对Y79细胞增殖的影响。结论 miR-30-5p通过下调FOXG1表达抑制视网膜母细胞瘤Y79细胞的增殖。

关键词: 视网膜母细胞瘤, miR-30-5p, FOXG1, 细胞增殖, 人视网膜内皮细胞

Abstract: Objective To investigate the effect of microRNA-30-5p(miR-30-5p)on the proliferation of retinoblastoma cells. Methods The expression of miR-30-5p in retinoblastoma cell lines and normal human retinal epithelial cells(HRECs)was determined using quantitative Real-time polymerase chain reaction(qRT-PCR). Bioinformatic analysis and prediction of miR-30-5p target genes were performed using the TargetScan database. Dual-luciferase reporter experiments verified the 3'-UTR binding ability and targeting relationship of miR-30-5p and FOXG1. The effect of miR-30-5p on the mRNA and protein expression of FOXG1 was determined using qRT-PCR and Western blotting, respectively. The effects of the miR-30-5p-FOXG1 pathway on cell proliferation were examined in Y79 cells by transient transfection. Results miR-30-5p expression was decreased and FOXG1 expression was increased in retinoblastoma cells compared with that in HRECs. Bioinformatics prediction results revealed the binding sites of miR-30-5p and FOXG1. qRT-PCR and western blot results showed that miR-30-5p negatively regulated FOXG1 mRNA and protein expression. Dual-luciferase assay confirmed a targeting relationship between miR-30-5p and FOXG1 3'-UTR. Transient transfection of miR-30-5p increased the expression level of miR-30-5p in Y79 cells. Overexpression of FOXG1 reversed the inhibition of Y79 cell proliferation. Conclusion miR-30-5p inhibits the proliferation of Y79 retinoblastoma cells by downregulating FOXG1 expression.

Key words: Retinoblastoma, Y79, miR-30-5p, FOXG1, Cell proliferation, Human retinal epithelial cells

中图分类号:

R774.1

颜繁诚,蒋现,柴一杰,王昊森,孟照洋,汪晓磊,王艳玲,冯雪. miR-30-5p通过下调FOXG1表达抑制视网膜母细胞瘤细胞增殖[J]. 山东大学耳鼻喉眼学报, 2022, 36(5): 63-69.

YAN Fancheng, JIANG Xian, CHAI Yijie, WANG Haosen, MENG Zhaoyang, WANG Xiaolei, WANG Yanling,. miR-30-5p inhibits retinoblastoma cell proliferation by downregulating FOXG1 expression[J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2022, 36(5): 63-69.

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