锌指蛋白331的表达在肝癌中受启动子区甲基化调控 杨志华 郭明洲 毛 .docVIP

锌指蛋白331的表达在肝癌中受启动子区甲基化调控 杨志华 郭明洲 毛高平 [摘要] 目的 探讨锌指蛋白331（ZNF331）启动子区甲基化在肝癌发生发展中的作用。方法 采用甲基化特异性PCR（MSP）ZNF331在肝癌细胞系和50例原发性肝癌组织中的甲基化状况，同时采用半定量RT-PCR分析5-杂氮-2’-脱氧胞苷（5-Aza）ZNF331的表达情况。结果 在6株肝癌细胞系中SW1990表达缺失、HBXF344、PLC/PRF/5、HepG2、BEL-7402低表达， SNU449表达正常。5-Aza处理后SW1990、HBXF344、HepG2、BEL-7402表达恢复正常。50例原发性肝癌标本中，ZNF331启动子区甲基化率为80%（40/50），而10例正常肝脏组织中ZNF331启动子区均无甲基化。ZNF331启动子区甲基化与肝癌患者的性别、年龄、肿瘤直径大小、血清甲胎蛋白值、肝炎病毒感染、分化水平及TNM分期均无关联(p0.05)。结论ZNF331启动子区高甲基化是肝癌的频发事件，ZNF331基因表达受启动子区甲基化调控。 [关键词] 肝癌； DNA甲基化； ZNF331基因； 甲基化特异性PCR Zinc-finger protein 331 is regulated by promoter region hypermethylation in human hepatocellular carcinoma YANG Zhi-hua，GUO Ming-zhou，MAO Gao-ping. Auhui Medical University，Hefei 230032，China Corresponding author：MAO Gao-ping Email: maogao ping@ [Abstract] Objective To analyze the methylation status of zinc-finger protein 331 (ZNF331) and the regulation of ZNF331 expression in human hepatic cancer. Methods Methylation specific PCR (MSP) and semi-quantitative RT-PCR were employed to detect ZNF331 promoter region methylation and the expression. Six human hepatic cancer cell lines and 50 cases of human primary hepatic cancer were included in this study. Results ZNF331 promoter region complete methylation was found in SW1990 cell line and partial methylation was detected in HBXF344、PLC/PRF/5、HepG2、BEL-7402 cell lines. Unmethylation was found in SNU449 cell line. Loss of expression was detected in SW1990 cell line and weak expression was found in HBXF344、PLC/PRF/5、HepG2、BEL-7402. In SNU449 cell line, ZNF331 was expressed. Restoration of ZNF331 expression was detected in SW1990 cell line and increased expression was found in HBXF344、HepG2、BEL-7402 cell lines after 5-Aza treatment. No expression changes were examined in SNU449 and PLC/PRF/5 cell lines before and after 5-Aza treatment. Promoter region hypermethylation was found in 80% primary human hepatic cancer (40/50). But no methylation was detected in normal liver tissues (0/10). No correlation was found b