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| MiR-122通过靶向SIRT1抑制肝癌细胞的增殖、迁移和侵袭 |
| MiR-122 inhibits the proliferation, migration and invasion of liver cancer cells by targeting SIRT1 |
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| DOI：10.3969/j.issn.1007-8134.2023.04.05 |
| 中文关键词: 微小RNA-122 沉默信息调节因子1 肝癌 HepG2 增殖 迁移 侵袭 细胞周期蛋白D1 E-钙粘蛋白 |
| 英文关键词: microRNA-122 silent information regulator 1 liver cancer HepG2 proliferation migration invasion CyclinD1 E-cadherin |
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| 中文摘要: |
| 目的?探讨微小RNA-122（microRNA-122, miR-122）通过靶向沉默信息调节因子1（silent information regulator 1, SIRT1）对肝癌细胞增殖、迁移和侵袭的影响。方法?用实时荧光定量PCR（real-time quantitative PCR, RT-qPCR）法检测肝癌组织和细胞中miR-122和SIRT1 mRNA表达水平；用生物信息学和双荧光素酶报告基因分析并验证miR-122和SIRT1靶向关系。培养肝癌细胞HepG2，将其分为空白对照组（control组）、miR-122过表达阴性对照组（miR-NC mimic组）、miR-122过表达组（miR-122 mimic组）、miR-122过表达和SIRT1过表达阴性对照组（miR-122 mimic+pcDNA组）、miR-122过表达和SIRT1过表达组（miR-122 mimic+pcDNA-SIRT1组）。用细胞计数试剂（cell counting kit-8, CCK-8）和5-乙炔基-2’脱氧尿嘧啶核苷分析各组细胞增殖情况；划痕愈合实验和Transwell实验分析各组细胞迁移和侵袭能力；用Western blot法分析各组细胞中增殖、迁移和侵袭相关蛋白p53、细胞周期蛋白D1、E-钙粘蛋白、N-钙粘蛋白和波形蛋白及SIRT1蛋白表达水平。结果?MiR-122表达水平在肝癌组织和细胞中下调，SIRT1 mRNA表达水平在肝癌组织和细胞中上调（P＜0.05）。MiR-122负靶向调控SIRT1表达（P＜0.05）。过表达miR-122抑制HepG2细胞增殖、迁移和侵袭，降低SIRT1、细胞周期蛋白D1、N-钙粘蛋白和波形蛋白蛋白水平，升高p53和E-钙粘蛋白蛋白水平（P＜0.05）。然而，SIRT1过表达可部分逆转过表达miR-122对HepG2细胞的作用（P＜0.05）。结论?MiR-122通过负靶向调控SIRT1抑制肝癌细胞增殖、迁移和侵袭。 |
| 英文摘要: |
| Objective To explore the effects of microRNA-122 (miR-122) on the proliferation, migration and invasion of liver cancer cells by targeting silent information regulator 1 (SIRT1). Methods?Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of miR-122 and SIRT1 mRNA in liver cancer tissues and cells; bioinformatics and dual luciferase reporter gene were used to analyze and verify the targeting relationship between miR-122 and SIRT1. The liver cancer cells HepG2 were cultured and divided into blank control group (control group), miR-122 overexpression negative control group (miR-NC mimic group), miR-122 overexpression group (miR-122 mimic group), miR-122 overexpression and SIRT1 overexpression negative control group (miR-122 mimic+pcDNA group), and miR-122 overexpression and SIRT1 overexpression group (miR-122 mimic+pcDNA-SIRT1 group). Cell counting kit-8 (CCK-8) and 5-Ethynyl-2’ -deoxyuridine (EdU) methods were used to analyze cell proliferation in each group; scratch healing test and Transwell test were used to analyze cell migration and invasion abilities of each group; Western blot was used to analyze the expression levels of proliferation, migration and invasion related proteins of cells in each group, including p53, CyclinD1, E-cadherin, N-cadherin, vimentin and SIRT1 proteins. Results?The expression level of miR-122 was down-regulated in liver cancer tissues and cells, and the expression level of SIRT1 mRNA was up-regulated in liver cancer tissues and cells (P＜0.05). MiR-122 negatively targeted the expression of SIRT1 (P＜0.05). Overexpression of miR-122 inhibited HepG2 cell proliferation, migration and invasion, decreased SIRT1, CyclinD1, N-cadherin and vimentin protein levels, and increased p53 and E-cadherin protein levels (P＜0.05). However, overexpression of SIRT1 could partially reverse the effect of overexpression of miR-122 on HepG2 cells (P＜0.05). Conclusion?MiR-122 inhibits the proliferation, migration and invasion of liver cancer cells by negatively targetingly regulating SIRT1. |
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