黑色素瘤中HOXA9启动子区甲基化水平与基因表达的关联及其对患者预后的影响  

Association of HOXA9 promoter methylation and gene expression and its effect on prognosis in melanoma

作　　者：李婷婷[1] 王鹏[1] 赵娟[1] 康晓静[1] Li Tingting;Wang Peng;Zhao Juan;Kang Xiaojing(Department of Dermatology and Venereology,People’s Hospital of Xinjiang Uygur Autonomous Region,Xinjiang Clinical Research Center for Dermatology and Venereology,Xinjiang Key Laboratory of Dermatology Research,Wulumuqi 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院皮肤性病科,新疆皮肤性病临床医学研究中心,新疆皮肤病研究重点实验室,新疆乌鲁木齐830001

出　　处：《实用肿瘤杂志》2025年第1期53-61,共9页Journal of Practical Oncology

基　　金：新疆维吾尔自治区自然科学基金(2021D01C201);新疆维吾尔自治区人民医院院内项目(20200104)。

摘　　要：目的分析同源盒A9(homeobox A9,HOXA9)启动子区甲基化水平与基因表达的相关性并探索HOXA9启动子区甲基化水平与黑色素瘤发生和预后的关系。方法采用MethSurv数据库分析人皮肤黑色素瘤(skin cutaneous melanoma,SKCM)中HOXA9基因启动子区甲基化水平。采用基因表达谱交互分析(Gene Expression Profiling Interactive Analysis,GEPIA)数据库分析SKCM组织和正常皮肤组织中HOXA9 mRNA表达水平。DNA甲基化交互式可视化数据库(DNA Methylation Interactive Visualization Database,DNMIVD)分析SKCM中HOXA9启动子区甲基化水平与基因表达的关系。人类蛋白质图谱(Human Protein Atlas,HPA)数据库分析SKCM组织和正常皮肤组织中HOXA9蛋白表达水平。在人黑色素瘤A375细胞中进行验证:20μmol/L 5-氮杂胞苷(5-azacytidine,5-azaC)作用于体外培养的A375细胞72 h,以未经药物干预的常规培养A375细胞作为对照组;采用甲基化特异性聚合酶链式反应(polymerase chain reaction,PCR)检测HOXA9基因启动子区甲基化状态;采用定量反转录PCR(quantitative reverse transcriptase-PCR,qRT-PCR)和蛋白质印迹法检测HOXA9 mRNA和蛋白表达水平。SurvivalMeth数据库分析SKCM中HOXA9启动子区甲基化水平与患者总生存期(overall survival,OS)的关系。结果MethSurv数据库分析表明,SKCM中HOXA9基因甲基化多发生在启动子区CpG岛,且多为高甲基化水平。GEPIA数据库分析表明,SKCM组织中HOXA9 mRNA表达水平低于正常皮肤组织(P<0.01),不同分期的SKCM患者HOXA9 mRNA表达水平比较,差异无统计学意义(P>0.05)。DNMIVD数据库分析表明,SKCM中HOXA9基因启动子区甲基化水平与其基因表达呈负相关(r=-0.34,P<0.01)。HPA数据库分析显示,HOXA9在正常皮肤组织内中等表达,在SKCM组织内低表达。A375细胞实验显示,对照组HOXA9基因启动子区表现为甲基化状态,经5-azaC处理72 h后,HOXA9基因启动子区甲基化与非甲基化状态并存,HOXA9 mRNA和蛋白表达量均较Objective To explore the correlation between the promoter methylation and gene expression of homeobox A9(HOXA9),and the association between the methylation of HOXA9 promoter and the occurrence and prognosis of melanoma.Methods The MethSurv database was used to analyze HOXA9 promoter methylation levels in human skin cutaneous melanoma(SKCM).The Gene Expression Profiling Interactive Analysis(GEPIA)database was used to analyze HOXA9 mRNA expression in SKCM and normal skin tissues.The DNA Methylation Interactive Visualization Database(DNMIVD)was used to analyze the relationship between HOXA9 promoter methylation levels and gene expression in SKCM.The Human Protein Atlas(HPA)database was used to analyze HOXA9 protein expression in SKCM and normal skin tissues.Human melanoma A375 cells were used to validate the results.20μmol/L 5-azacytidine(5-azaC)was applied to cultured A375 cells for 72 h,and A375 cells without drug intervention were used as a control group.Methylation-specific polymerase chain reaction(PCR)was used to detect the methylation status of HOXA9 promoter.The mRNA and protein levels of HOXA9 in A375 cells were detected by quantitative reverse transcriptase-PCR(qRT-PCR)and Western blotting.The SurvivalMeth database was used to analyze the relationship between the methylation level of HOXA9 promoter region and the overall survival(OS)of melanoma patients.Results The MethSurv database analysis indicated that HOXA9 gene methylation in SKCM occurred mostly in CpG islands of the promoter region,and the methylation levels were mostly high.The GEPIA database analysis showed that the expression level of HOXA9 mRNA in SKCM tissues was significantly lower than that in normal skin tissues(P<0.01).However,no statistically significant difference was observed when comparing the HOXA9 mRNA expression levels of melanoma patients with different stages(P>0.05).The DNMIVD database analysis showed that the promoter methylation level of HOXA9 in SKCM was negatively correlated with its gene expression(r=-0.34,P<0.01).The HPA d

关 键 词：黑色素瘤 同源盒A9 DNA甲基化 数据挖掘 A375细胞 

分 类 号：R739.5[医药卫生—肿瘤]