遵义医科大学学报2024，Vol.47Issue(2) ：135-144.

N6-甲基腺苷(m6A)转移酶METTL3和m6A调控LINC01465在肝细胞癌增殖转移中的作用机制

Mechanism of N6-methyladenosine(m6A)transferase METTL3 and m6A reg-ulation of LINC01465 in the proliferation and metastasis of hepatocellular car-cinoma

刘肸琳 龙朴泽 黄云美 罗春英
遵义医科大学学报2024，Vol.47Issue(2) ：135-144.
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N6-甲基腺苷(m6A)转移酶METTL3和m6A调控LINC01465在肝细胞癌增殖转移中的作用机制

Mechanism of N6-methyladenosine(m6A)transferase METTL3 and m6A reg-ulation of LINC01465 in the proliferation and metastasis of hepatocellular car-cinoma

刘肸琳 1龙朴泽 2黄云美 3罗春英4
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作者信息

  • 1. 右江民族医学院研究生学院,广西百色 533000
  • 2. 湖北医药学院附属东风医院病理科,湖北十堰 442008
  • 3. 右江民族医学院附属医院病理科,广西百色 533000
  • 4. 右江民族医学院附属医院病理科,广西百色 533000;广西肝胆疾病分子病理重点实验室,广西百色 533000
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摘要

目的 探讨甲基转移酶样3(METTL3)在肝癌细胞中调控m6A水平,影响长链非编码(lncRNA)LINC01465的表达,促进肝癌细胞增殖转移的机制.方法 利用人类癌症基因组图谱(TCGA)数据库分析METTL3在肝癌组织和正常肝组织中的表达差异及与患者预后的关系;用Western blot、RT-qPCR分析验证METTL3在细胞、组织中的表达;用m6A比色法验证肝癌细胞及正常肝细胞中的m6A水平;敲低METTL3后进行CCK-8、克隆形成、Transwell迁移、侵袭实验,研究METTL3对肝癌增殖转移的调控作用;转录组RNA m6A甲基化测序确定METTL3的下游调控基因LINC01465.RT-qPCR验证沉默METTL3后LINC01465的表达量,以及LINC01465在肝细胞癌中的表达水平.结果 METTL3在肝癌组织和细胞中明显高表达且与患者预后较差相关;高表达的METTL3促进HCC细胞增殖、迁移和侵袭;METTL3影响LINC01465的m6A水平及其表达量来促进肝细胞癌的发展进程.结论 METTL3可通过m6A依赖的方式来影响LINC01465的表达水平,促进肝癌细胞增殖转移,METTL3可能成为肝癌预后及治疗的靶向标志物.

Abstract

Objective To investigate the mechanism of methyltransferase like 3(METTL3)in hepatocellular carcinoma cells to regulate the level of m6 A,affect the expression of long-chain non-coding(lncRNA)LINC01465,and promote the proliferation and metastasis of hepatocellular carcinoma cells.Methods The Canc-er Genome Atlas(TCGA)database was used to analyze the expression differences of METTL3 in hepatocellular carcinoma tissues and normal liver tissues and explain its relationship with patients'prognosis;the expression of METTL3 in cells and tissues was verified by western blot and RT-qPCR analyses;the levels of m6A were verified by the m6A colorimetric assay in hepatocellular carcinoma cells and normal hepatocytes;and after the knock-down of METTL3,the CCK-8,clone formation,Transwell migration,and invasion assays was used to study the regulatory role of METTL3 on the proliferation and metastasis of hepatocellular carcinoma;transcriptome RNA m6A methylation sequencing was performed to determine LINC01465,the downstream regulatory gene of MET-TL3.RT-qPCR was performed to verify the expression of LINC01465 after silencing of METTL3 and the expres-sion level of LINC01465 in hepatocellular carcinoma.Results METTL3 was significantly highly expressed in hep-atocellular carcinoma tissues and cells and was associated with poorer prognosis of patients;highly expressed METTL3 promoted the proliferation,migration and invasion of HCC cells;METTL3 affected the m6A level of LINC01465 and its expression,promoting the developmental process of hepatocellular carcinoma.Conclusion METTL3 can affect the expression level of LINC01465 in an m6A-dependent manner to promote the proliferation and metastasis ability of hepatocellular carcinoma cells,suggesting that METTL3 may become a target marker for the prognosis and treatment of hepatocellular carcinoma.

关键词

甲基转移酶样3/m6A甲基化/LINC01465/肝细胞癌

Key words

methyltransferase like 3/N6-methyladenosine methylation/LINC01465/hepatocellular carcinoma

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基金项目

广西自然科学基金资助项目(2017GXNSFGA198002)

百色市区域多发病联合专项计划项目(20224104)

出版年

2024
遵义医科大学学报
遵义医科大学

遵义医科大学学报

CSTPCD
ISSN：2096-8159
参考文献量33
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