肝细胞癌中与m^(6)A相关的lncRNAs挖掘及DACH1在其预后的免疫特性  

Exploration of m^(6)A-Related lncRNAs and the Role of DACH1 in Prognosis and Immune Characteristics of Hepatocellular Carcinoma

作　　者：刘虹汝 张洪源 张紫栋 邹静 舒林丹 刘威东 刘衡 张成桂 张钰哲 LIU Hongru;ZHANG Hongyuan;ZHANG Zidong;ZOU Jing;SHU Lindan;LIU Weidong;LIU Heng;ZHANG Chenggui;ZHANG Yuzhe(College of Basic Medical Science,Dali University,Dali 671000,Yunnan,China;Gansu Health Vocational College,Lanzhou 730207,China;Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D,Dali University,Dali 671000,Yunnan,China;Department of Respiratory Medicine,The First Affiliated Hospital of Dali University,Dali 671000,Yunnan,China)

机构地区：[1]大理大学基础医学院,云南大理671000 [2]甘肃卫生职业学院,甘肃兰州730207 [3]云南省昆虫医药研发重点实验室,云南大理671000 [4]大理大学第一附属医院呼吸内科,云南大理671000

出　　处：《昆明理工大学学报（自然科学版）》2024年第5期119-132,共14页Journal of Kunming University of Science and Technology(Natural Science)

基　　金：云南省地方本科高校(部分)基础研究联合专项面上项目(202001BA070001-064,202101BA070001-102);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).

摘　　要：探讨N6-甲基腺嘌呤(N6-methyladenosine,m^(6)A)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(Hepatocellular 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m^(6)A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m^(6)A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m^(6)A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m^(6)A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.This study aimed to investigate the impact of N6-methyladenosine(m^(6)A)-related long non-coding RNAs(LncRNAs)and Dachshund homolog 1(DACH1)on the prognosis and immune infiltration of hepatocellular carcinoma(HCC).Using bioinformatics,we constructed a risk model involving DACH1 and m^(6)A-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 m^(6)A-related LncRNAs were involved in the progression of HCC.Through univariate,multivariate,and LASSO regression analyses,11 m^(6)A-related 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 m^(6)A-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 diagnosis and prognosis of HCC.

关 键 词：肝细胞癌 m^(6)A 长链非编码RNA DACH1 免疫微环境 

分 类 号：R735.7[医药卫生—肿瘤]