microRNA-21靶向调控Wnt2基因对肝癌细胞HepG2增殖与迁移的影响

王泽鑫; 关利君; 李建明; 王志超; 薛梦若; 秦孝军

doi:10.3969/j.issn.1001-5256.2020.04.019

microRNA-21靶向调控Wnt2基因对肝癌细胞HepG2增殖与迁移的影响

DOI: 10.3969/j.issn.1001-5256.2020.04.019

内蒙古医科大学附属医院介入放射科

基金项目:

内蒙古自治区自然科学基金(2017MS0890)；

详细信息

中图分类号: R735.7
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出版历程
- 收稿日期: 2019-11-15
- 出版日期: 2020-04-20

Effect of targeted regulation of the Wnt2 gene by microRNA( microRNA-21) on the proliferation and migration of HepG2 hepatoma cells

Department of Interventional Radiology,The Affiliated Hospital of Inner Mongolia Medical University

Research funding:

摘要

摘要: 目的探讨分析microRNA-21（miR-21）靶向调控Wnt2基因对肝癌细胞HepG2增殖与迁移的影响。方法采用实时荧光定量PCR法检测肝癌细胞HepG2及正常肝细胞株LO2 miR-21的表达情况。对HepG2细胞转染miR-21抑制剂,分析转染后抑制剂组与对照组miR-21的表达情况、细胞增殖、迁移及凋亡情况,检测两组细胞Wnt2蛋白表达水平,并采用双荧光素酶报告基因实验验证miR-21与Wnt2基因的关系。计量资料两组间比较采用t检验。结果 HepG2细胞miR-21相对表达水平明显高于LO2细胞（1. 978±0. 035 vs 1. 586±0. 022,t=16. 424,P <0. 05）。转染miR-21抑制剂后,抑制剂组miR-21相对表达水平较对照组显著降低（0. 857±0. 017 vs 1. 684±0. 039,t=33. 669,P <0. 05）。转染miR-21抑制剂24、48、72 h后,抑制剂组HepG2细胞增殖能力均较对照组显著降低（P值均<0. 05）;抑制剂组穿过Tranwell小室的细胞数显著低于对照组（83. 72±...
- 微RNAs /
- Wnt2蛋白质 /
- 癌,肝细胞 /
- 细胞增殖 /
- 细胞运动
Abstract: Objective To investigate the effect of targeted regulation of the Wnt2 gene by microRNA( miR-21) on the proliferation and migration of HepG2 hepatoma cells. Methods Quantitative real-time PCR was used to measure the mRNA expression of miR-21 in HepG2 hepatoma cells and normal liver cell line LO2. HepG2 cells were transfected with miR-21 inhibitor,and then the expression of miR-21 and cell proliferation,migration,and apoptosis were analyzed for the inhibitor group and the control group. The protein expression of Wnt2 was measured for the two groups,and dual-luciferase reporter assay was used to verify the association between miR-21 and the Wnt2 gene. The t-test was used for comparison of continuous data between groups. Results The relative expression of miR-21 in HepG2 cells was significantly higher than that in LO2 cells( 1. 978 ± 0. 035 vs 1. 586 ± 0. 022,t = 16. 424,P < 0. 05). After the transfection of miR-21 inhibitor,the inhibitor group had significantly lower expression of miR-21 than the control group( 0. 857 ± 0. 017 vs 1. 684 ± 0. 039,t= 33. 669,P < 0. 05). Compared with the control group after the transfection of miR-21 inhibitor,the inhibitor group had a significant reduction in the proliferation ability of HepG2 cells( P < 0. 05),a significantly lower number of cells passing through the Transwell chamber( 83. 72 ± 15. 06 vs 147. 85 ± 20. 64,t = 4. 347,P < 0. 05),and a significantly higher cell apoptosis rate( 25. 67% ± 3. 95% vs 10. 27%± 2. 14%,t = 5. 937,P < 0. 05). The inhibitor group had significantly lower relative expression of Wnt2 in HepG2 cells than the control group( 0. 862 ± 0. 127 vs 1. 306 ± 0. 218,t = 3. 048,P < 0. 05). TargetScan software showed that miR-21 inhibitor significantly inhibited the luciferase activity of the cells transfected with wild-type Wnt2-3'UTR plasmid( 0. 972 ± 0. 102 vs 0. 612 ± 0. 092,t = 4. 219,P <0. 05),while there was no effect on the luciferase activity of the cells transfected with mutant Wnt2-3' UTR plasmid( 0. 982 ± 0. 093 vs0. 911 ± 0. 128,t = 0. 972,P > 0. 05). Conclusion Inhibition of miR-21 expression can effectively inhibit the proliferation and migration of HepG2 cells,promote the apoptosis of HepG2 cells,and inhibit the over-activation of the Wnt signaling pathway,and therefore,it may become one of the potential target genes for liver cancer treatment.
- microRNAs /
- Wnt2 protein /
- carcinoma,hepatocellular /
- cell proliferation /
- cell movement

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