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miR-222通过调控BCL2L13基因促进HBx-HepG2细胞生长

miR-222 enhances HBx-HepG2 cell growth via regulation of BCL2L13 gene
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摘要 目的:探讨HBx-Hep G2细胞中微小RNA-222(mi R-222)对BCL2L13基因表达的调控作用以及对细胞生长和凋亡的影响,并研究其潜在的分子作用机制。方法:利用实时荧光定量PCR检测mi R-222的表达水平;MTT和集落形成实验检测细胞的生长;流式细胞术检测细胞周期和凋亡;构建BCL2L13 3’UTR双萤光素酶报告载体,通过采用双萤光素酶报告实验验证mi R-222的靶基因。结果:与正常的肝细胞L02相比,mi R-222在HBx-Hep G2细胞过量表达(P<0.05)。mi R-222过表达可促进HBx-Hep G2细胞的生长,改变细胞周期,降低细胞的凋亡率;mi R-222表达下调可抑制HBx-Hep G2细胞的生长,改变细胞周期,增加细胞的凋亡率,和对照组相比差异有统计学显著性(P<0.05)。与正常肝细胞L02相比,BCL2L13在HBx-Hep G2细胞表达下调(P<0.05);mi R-222表达下调可促进BCL2L13的表达(P<0.05)。双萤光素酶报告实验和pc DNA3.1-BCL2L13转染实验结果提示mi R-222可以通过作用于BCL2L13的3’UTR区,负向调控其表达,从而促进细胞的生长。结论:mi R-222可以通过靶向调控BCL2L13基因进而促进HBx-HepG2细胞的生长。 AIM: To investigate the regulation of mi R-222 on BCL2L13 gene and its effect on the growth and apoptosis of HBx-Hep G2 cells,and to explore the underlying molecular mechanisms. METHODS: The expression level of mi R-222 was detected by RT-q PCR. The HBx-Hep G2 cell growth was examined by MTT and colony formation assays. The cell cycle and apoptosis were analyzed by flow cytometry. The recombination vector pmir GLO-BCL2L13 was constructed,and dual-luciferase reporter experiment was performed to validate the target of mi R-222. RESULTS: The expression level of mi R-222 in the HBx-Hep G2 cells was significantly higher than that in the L02 cells( P 0. 05). Over-expression of mi R-222 enhanced HBx-Hep G2 cell growth,changed cell cycle,and inhibited apoptosis( P 0. 05). Knockdown of mi R-222 reduced HBx-Hep G2 cell growth,changed cell cycle,and increased cell apoptotic rate( P 0. 05). BCL2L13 was down-regulated in the HBx-Hep G2 cells as compared with L02 cells( P 0. 05),and knockdown of mi R-222 in the HBxHep G2 cells increased the expression level of BCL2L13( P 0. 05). The results of dual-luciferase reporter assay and restore experiment showed that mi R-222 negatively regulated the expression of BCL2L13 via targeting 3'UTR of BCL2L13,resulting in the promotion of HBx-Hep G2 cell growth. CONCLUSION: mi R-222 enhances HBx-Hep G2 cell growth via down-regulation of BCL2L13.
出处 《中国病理生理杂志》 CAS CSCD 北大核心 2016年第8期1389-1394,共6页 Chinese Journal of Pathophysiology
基金 广东省科技计划项目(No.20130319c)
关键词 微小RNA-222 HBx-HepG2细胞 细胞生长 细胞周期 细胞凋亡 BCL2L13基因 Micro RNA-222 HBx-Hep G2 cells Cell growth Cell cycle Apoptosis BCL2L13 gene
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