摘要
探讨N6-甲基腺嘌呤(N6-methyladenosine,m6A)修饰相关长链非编码RNA(Long non-coding RNA,LncRNA)及细胞命运决定因子 1(Dachshund homolog1,DACH1)对肝细胞癌(Hepato-cellular carcinoma,HCC)预后及免疫浸润的影响.运用生物信息学方法构建DACH1与m6A相关LncRNAs的风险模型、进行了聚类分析、免疫浸润和肿瘤微环境(Tumor micro-environment,TME)分析、评估了风险评分及临床相关性.研究结果表明,有56个与m6A相关的LncRNAs参与了肝细胞癌(Hepatocellular Carcinoma,HCC)的进展,通过单、多因素及LASSO回归分析,最终筛选出11个与DACH1表达最相关的m6A相关联LncRNAs,包括:AC027097.1、LINC01138、LINC01224、DDX11-AS1、MKLN1-AS、AL031985.3、AC098484.1、AL158166.1、POLH-AS1、WAC-AS1和NRAV,这些LncRNAs与免疫细胞浸润有密切相关;进一步的RT-qPCR、WB和IHC实验结果显示,HCC细胞中DACH1的mRNA表达显著低于正常肝细胞;而正常肝细胞和肝组织中的DACH1蛋白表达水平明显高于HCC细胞;此外,用5-aza-2dc(甲基化抑制剂)处理HCC细胞后,细胞迁移距离显著缩短,提示DACH1去甲基化后,可能影响其抑癌功能.本研究通过生物信息学分析和实验验证发现了 11个与DACH1表达相关的m6A修饰LncRNAs,并且DACH1在HCC发生、发展及免疫浸润中扮演了关键的抑癌角色,为HCC的早期诊断及临床预后提供了理论依据.
Abstract
This study aimed to investigate the impact of N6-methyladenosine(m6A)-related long non-coding RNAs(LncRNAs)and Dachshund homolog 1(DACH1)on the prognosis and immune infiltration of hepatocel-lular carcinoma(HCC).Using bioinformatics,we constructed a risk model involving DACH1 and m6A-related LncRNAs,conducted clustering analysis,immune infiltration,and tumor microenvironment(TME)analysis,and evaluated the risk scores and clinical correlations.The results showed that 56 m6A-related LncRNAs were in-volved in the progression of HCC.Through univariate,multivariate,and LASSO regression analyses,11 m6A-re-lated LncRNAs most correlated with DACH1 expression were identified,including AC027097.1,LINC01138,LINC01224,DDX11-AS1,MKLN1-AS,AL031985.3,AC098484.1,AL158166.1,POLH-AS1,WAC-AS1,and NRAV.These LncRNAs were closely associated with immune cell infiltration.Further RT-qPCR,Western blot(WB),and immunohistochemistry(IHC)experiments revealed that DACH1 mRNA expression in HCC cells was significantly lower than that in normal hepatocytes,and the DACH1 protein expression in normal liver cells and tissues was notably higher than in HCC cells.Additionally,treatment of HCC cells with 5-aza-2dc(a methylation inhibitor)significantly reduced cell migration distance,suggesting that DACH1 demethylation might enhance its tumor-suppressing function.Through bioinformatics analysis and experimental validation,this study identified 11 m6A-modified LncRNAs associated with DACH1 expression,highlighting the crucial role of DACH1 in HCC progression,development,and immune infiltration,providing theoretical support for the early di-agnosis and prognosis of HCC.
基金项目
云南省地方本科高校(部分)基础研究联合专项面上项目(202001BA070001-064)
云南省地方本科高校(部分)基础研究联合专项面上项目(202101BA070001-102)
云南省昆虫生物医药研发重点实验室开放课题(AG2024002)
大理大学博士科研启动基金项目(KYBS2018012)
万方数据