RNF180基因甲基化是胰腺癌的潜在诊断标志物  

Methylation of RNF180 gene is a potential diagnostic marker for pancreatic cancer

作　　者：李昕 段一梦 张美英 郭明洲 LI Xin;DUAN Yimeng;ZHANG Meiying;GUO Mingzhou(Basic Medical College,Zhengzhou University,Zhengzhou 450001;Department of Gastroenterology&Hepatology,the First Medical Center,Chinese PLA General Hospital,China)

机构地区：[1]郑州大学基础医学院,河南郑州450001 [2]中国人民解放军总医院第一医学中心消化内科医学部

出　　处：《胃肠病学和肝病学杂志》2022年第6期629-633,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家重点研发计划(2018YFA0208902,2020YFC2002705);国家自然科学基金委员会资助项目(81672318,U1604281);北京市自然科学基金重点项目(7171008);国家重点科学仪器专项计划(2011YQ03013405)。

摘　　要：目的探讨E3泛素连接酶家族的环指蛋白180(ring finger protein 180,RNF180)基因在胰腺癌中的甲基化情况及其表达调控机制。方法9个胰腺癌细胞系和112例原发性胰腺癌组织,以及半定量RT-PCR和甲基化特异性PCR(methylation specific PCR,MSP)技术被应用于RNF180基因的甲基化状态和表达调控研究。结果RNF180在CFPAC1、Panc5.04和PANC1细胞中高表达,MSP结果显示其启动子区域呈非甲基化状态;在T3M4和MIAPaCa2细胞中表达减少,其启动子区呈部分甲基化状态;在SW1990、Panc10.05、PANC3.11和AsPC1细胞中RNF180表达缺失,其启动子区呈完全甲基化状态。应用去甲基化药物5-aza-dc处理后,CFPAC1、Panc5.04和PANC1细胞中表达无变化,T3M4和MIAPaCa2细胞中表达增加,SW1990、Panc10.05、PANC3.11和AsPC1细胞中恢复表达。结果表明RNF180的表达受启动子区甲基化调控。RNF180基因在原发性胰腺癌中的甲基化率是31.25%(35/112)。RNF180甲基化与肿瘤分期有相关性(P<0.05),与年龄、性别、肿瘤大小、肿瘤位置、吸烟、饮酒等因素均无相关性(P均>0.05)。结论RNF180在胰腺癌中频繁发生甲基化,其表达受启动子区甲基化调节。RNF180甲基化是胰腺癌潜在的诊断和预后标志物。Objective To investigate the methylation status and expression regulation mechanism of E3 ubiquitin ligase family of ring finger protein 180(RNF180)gene in pancreatic cancer.Methods Nine pancreatic cancer cell lines(CFPAC1,Panc5.04,PANC1,T3M4,MIAPaCa2,SW1990,Panc10.05,PANC3.11 and AsPC1)and 112 cases of pancreatic cancer samples,as well as semi-quantitative RT-PCR and methylation specific PCR(MSP)techniques were employed to explore the methylation status and the expression regulation of RNF180 gene in pancreatic cancer.Results RNF180 was highly expressed in CFPAC1,Panc5.04 and PANC1 cells,reduced expression in T3M4 and MIAPaCa2 cells,and loss of expression in SW1990,Panc10.05,PANC3.11 and AsPC1 cells.The promoter region of RNF180 was found unmethylation in CFPAC1,Panc5.04 and PANC1 cells,partial methylation in T3M4 and MIAPaCa2 cells,and complete methylation in SW1990,Panc10.05,PANC3.11 and AsPC1 cells.The methylation status was reversely correlated to RNF180 expression.The expression of RNF180 was restored in SW1990,Panc10.05,PANC3.11 and AsPC1 cells,increased expression was found in T3M4 and MIAPaCa2 cells,and no expression changes were found in CFPAC1,Panc5.04 and PANC1 cells after treatment with 5-aza-dc.The results indicated that the expression of RNF180 was regulated by promoter region methylation in pancreatic cancer cells.RNF180 was methylated in 31.25%(35/112)of primary pancreatic cancer.Methylation of RNF180 was associated with tumor stage(P<0.05),and not associated with age,gender,tumor size,tumor location,smoking and drinking(all P>0.05).Conclusion RNF180 is frequently methylated in pancreatic cancer,and its expression is regulated by promoter methylation.RNF180 methylation is a potential diagnostic and prognostic marker of pancreatic cancer.

关 键 词：RNF180 胰腺癌 DNA甲基化 甲基化特异性PCR 

分 类 号：R735.9[医药卫生—肿瘤]