中国医科大学学报2026，Vol.55Issue(5)：416-422,7.DOI:10.12007/j.issn.0258-4646.2026.05.006
METTL3介导Cry2 m6A甲基化修饰促进人肝细胞癌Huh-7细胞乐伐替尼耐药
METTL3-mediated Cry2 m6A methylation modification promotes lenvatinib resistance in human hepatocellular carcinoma Huh-7 cells
摘要
Abstract
Objective To investigate the effects of cryptochrome 2(Cry2)and its N6-methyladenosine(m6A)methylation modification on lenvatinib resistance in human hepatocellular carcinoma(HCC)cells.Methods Human HCC cell line Huh-7 and its lenvatinib-re-sistant cells(Huh-7/LR)were cultured in vitro.si-Cry2,oe-Cry2,si-methyltransferase-like protein 3(METTL3),oe-METTL3 interference negative control(si-NC),and empty overexpression vector(Vector)plasmids were transfected into Huh-7/LR cells.The control group con-sisted of Huh-7/LR cells that were not transfected with the plasmid.The drug sensitivity of each group was assessed using the MTT assay.The rate of apoptosis in each group was determined by flow cytometry.The methylation level of RNA m6A in the cells was detected using colorimetry.The m6A methylation levels of Cry2 in each group were analyzed using the methylation RNA immunoprecipitation method.The mRNA and protein expression levels of Cry2 and METTL3 in each group were measured by real-time quantitative PCR and Western blot-ting,respectively.Results When compared to Huh-7 cells,the sensitivity of Huh-7/LR cells to lenvatinib was found to be significantly decreased(P<0.05),resulting in a drug resistance index of 6.320.Simultaneously,there was a significant decrease seen in the mRNA and protein expression levels of Cry2 in these cells(P<0.01).Additionally,the methylation level of RNA m6A and protein expression level of METTL3 were significantly elevated(P<0.05).Overexpression of Cry2 significantly augmented the sensitivity of the Huh-7/LR cells to lenvatinib and facilitated apoptosis(all P<0.05).Silencing METTL3 notably attenuated the methylation of Cry2 m6A in Huh-7/LR cells(P<0.01),upregulated the mRNA and protein expression levels of Cry2,increased the sensitivity of Huh-7/LR cells to lenvatinib,and promoted apoptosis(all P<0.05).Conversely,overexpressing METTL3 exerted an opposing effect.However,silencing Cry2 effectively reversed the sensitizing effect of METTL3 silencing to lenvatinib in Huh-7/LR cells.Conclusion METTL3 facilitates the elevation of Cry2 m6A methylation levels,thus resulting in the downregulation of Cry2 expression and consequently promoting resistance to lenvatinib in human HCC cells.关键词
肝细胞癌/乐伐替尼/隐花色素蛋白2/N6-甲基腺苷甲基化/耐药Key words
hepatocellular carcinoma/lenvatinib/cryptochrome 2/N6-methyladenosine/drug resistance分类
医药卫生引用本文复制引用
张喜华,何前进,郝泉水,李珊,李秀芳..METTL3介导Cry2 m6A甲基化修饰促进人肝细胞癌Huh-7细胞乐伐替尼耐药[J].中国医科大学学报,2026,55(5):416-422,7.基金项目
湖北省卫生健康委科研项目(WJ2021M086) （WJ2021M086）
