[1]张虹,陈颖平,陈圣文.miR-124-3p靶向调控KLF6基因表达对H2O2诱导的人晶状体上皮细胞增殖和凋亡的影响[J].眼科新进展,2020,40(2):125-130.[doi:10.13389/j.cnki.rao.2020.0030]
　ZHANG Hong,CHEN Yingping,CHEN Shengwen.Effect of miR-124-3p targeted regulation of KLF6 genes on proliferation and apoptosis of H2O2-induced human lens epithelial cells[J].Recent Advances in Ophthalmology,2020,40(2):125-130.[doi:10.13389/j.cnki.rao.2020.0030]
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miR-124-3p靶向调控KLF6基因表达对H2O2诱导的人晶状体上皮细胞增殖和凋亡的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年2期
页码:
125-130
栏目:
实验研究
出版日期:
2020-02-05

文章信息/Info

Title:
Effect of miR-124-3p targeted regulation of KLF6 genes on proliferation and apoptosis of H2O2-induced human lens epithelial cells
作者:
张虹; 陈颖平; 陈圣文
570216　海南省海口市，海南医学院第二附属医院眼科
Author(s):
ZHANG Hong; CHEN Yingping; CHEN Shengwen
Department of Ophthalmology，the Second Affiliated Hospital of Hainan Medical University，Haikou 570216，Hainan Province，China
关键词:
miR-124-3p; KLF6; 人晶状体上皮细胞; 氧化应激; 增殖; 凋亡
Keywords:
miR-124-3p; KLF6; human lens epithelial cells; oxidative stress; proliferation; apoptosis
分类号:
R776
DOI:
10.13389/j.cnki.rao.2020.0030
文献标志码:
A
摘要:
目的　探讨微小RNA-124-3p（miR-124-3p）对H2O2诱导的人晶状体上皮细胞增殖及凋亡的影响及其靶向调控 Krüppel样因子6(Krüppel like factor 6，KLF6)的机制。方法　按HLE-B3细胞处理方式的不同，将其分为HLE-B3组、HLE-B3+ H2O2组、HLE-B3+H2O2+miR-NC组、HLE-B3+H2O2+miR-124-3p组、HLE-B3+H2O2+si-NC组、HLE-B3+H2O2+si-KLF6组、miR-124-3p+pcDNA3.1组、miR-124-3p+pcDNA3.1-KLF6组。利用MTT实验检测各组HLE-B3细胞增殖活性，流式细胞仪检测各组HLE-B3细胞凋亡。双荧光素酶报告基因实验验证miR-124-3p与KLF6的靶向关系。Western blot检测各组HLE-B3细胞中Cyclin D1、P21、Bax、Bcl-2蛋白表达。结果　H2O2可抑制人晶状体上皮HLE-B3细胞中miR-124-3p的表达（HLE-B3组0.79±0.07、HLE-B3+H2O2组0.31±0.03）（P＜0.05），而明显促进KLF6 的表达；miR-124-3p过表达或抑制KLF6表达可促进HLE-B3细胞增殖，抑制HLE-B3细胞凋亡（19.34±1.27、7.66±0.38；19.29±1.33、11.46±1.02），促进Cyclin D1（0.39±0.04、0.89±0.08；0.39±0.04、0.74±0.07）、Bcl-2表达（0.29±0.03、0.74±0.07；0.29±0.03、0.60±0.06），抑制P21（0.73±0.07、0.26±0.03；0.79±0.07、0.33±0.03）、Bax表达（0.86±0.08、0.37±0.03；0.86±0.08、0.51±0.05）。共转染miR-124-3p mimics与WT-KLF6可明显降低HLE-B3细胞的荧光素酶活性（0.27±0.03、0.98±0.08）（P＜0.05）；过表达KLF6可逆转miR-124-3p对H2O2诱导的人晶状体上皮细胞HLE-B3细胞增殖及凋亡的作用。结论　miR-124-3p可通过靶向调控 KLF6表达进而促进H2O2诱导的人晶状体上皮细胞HLE-B3细胞增殖并抑制其凋亡。
Abstract:
Objective　To investigate the effect of microRNA-124-3p (miR-124-3p) on proliferation and apoptosis of H2O2-induced human lens epithelial cells,and its mechanism of targeted regulation of Krüppel-like factor 6 (KLF6).Methods　According to different cell treatment methods,HLE-B3 cells were divided into HLE-B3 group,HLE-B3+H2O2 group,HLE-B3+H2O2+miR-NC group,HLE-B3+H2O2+miR-124-3p group,HLE-B3+H2O2+si-NC group,HLE-B3+H2O2+si- KLF6 group,miR-124-3p+pcDNA3.1 group and miR-124-3p+pcDNA3.1-KLF6 group.HLE-B3 cell proliferation activity was detected by MTT assay and HLE-B3 cell apoptosis was detected by flow cytometry.The targeting relationship between miR-124-3p and KLF6 was verified by dual luciferase reporter gene assay.Protein expression of Cyclin D1,P21,Bax and Bcl-2 in HLE-B3 cells was detected by Western blot.Results　H2O2 inhibited the expression of miR-124-3p in human lens epithelial HLE-B3 cells (HLE-B3 group:0.79±0.07;HLE-B3+H2O2 group:0.31±0.03) (P<0.05),and significantly promoted the expression of KLF6.Overexpression or inhibition of KLF6 expression by miR-124-3p promoted the proliferation of HLE-B3 cells and inhibited the apoptosis of HLE-B3 cells (19.34±1.27 vs 7.66±0.38;19.29±1.33 vs 11.46±1.02),and promoted the expression of Cyclin D1 (0.39±0.04 vs 0.89±0.08;0.39±0.04 vs 0.74±0.07),promoted the expression of Bcl-2 (0.29±0.03 vs 0.74±0.07;0.29±0.03 vs 0.60±0.06),inhibited the expression of P21 (0.73±0.07 vs 0.26±0.03;0.79±0.07 vs 0.33±0.03),inhibited the expression of Bax (0.86±0.08 vs 0.37±0.03;0.86±0.08 vs 0.51±0.05).Co-transfection of miR-124-3p mimics with WT-KLF6 significantly reduced luciferase activity in HLE-B3 cells (0.27±0.03 vs 0.98±0.08) (P<0.05).Overexpression of KLF6 reversed the effect of miR-124-3p on proliferation and apoptosis of H2O2-induced HLE-B3 cells.Conclusion　miR-124-3p can promote the proliferation of HLE-B3 cells induced by H2O2 and inhibit its apoptosis by targeting KLF6 expression.

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备注/Memo

备注/Memo:
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更新日期/Last Update: 2020-02-05