人肝癌组织GGT基因甲基化状态与酶蛋白异常表达关系的研究.docVIP

???? 人肝癌组织GGT基因甲基化状态与酶蛋白异常表达关系的研究 ???? ???? 摘要　目的 探讨肝癌发生过程中－谷氨酰转肽酶(GGT)的表达及其改变机制。 方法 将肝癌、癌旁及远癌组织总GGT和RNA纯化, 观察总GGT、膜结合和可溶性GGT的表达,并以逆转录巢式PCR扩增GGT非编码区基因片段, 分析M3位点的甲基化状态。 结果 肝癌、癌旁和远癌组总RNA浓度呈明显的梯度升高趋势(P0.05)；癌组织总GGT、膜结合和可溶性GGT均高于癌周和远癌组织(P0.05)；在癌、癌旁和远癌组织中,特定基因片段的扩增检出率分别为100%，85%和75%, M3位点低甲基化频率分别为75%，55%和50%。 结论 肝癌组织中GGT呈异常表达与基因低甲基化状态有关, 分析GGT基因可望成为监测肝细胞癌变的灵敏方法。关键词　癌，肝细胞 甲基化 GGT基因 表达 Relationship between methylation status of -glutamyl transpeptidase (GGT) genes and abnormal expression of its enzyme proteins in tissues of human hepatomas YAO Dengfu, TANG Qiyun, MENG Xianyong, et al. Affiliated Hospital, Nantong Medical College, Nantong 226001 Abstract　Objective To explore the mechanism of expression and alteration of -glutamyl transpeptidase (GGT) genes during the development of human hepatomas. Methods The total GGT protein and total RNA were purified in human hepatomas, adjacent paracancerous and distal cancerous tissues. The specific activities of total GGT, membrane-combine GGT and soluble GGT were investigated, the GGT gene of -NC region was amplified by using a nest RT-PCR assay, and methylation status of GGT gene M3 site were analyzed in the present study. Results An increasing tendency (P0.05) of total RNA concentrations was found from cancer in distal cancerous tissues; the specific activities (U/g) of total GGT, membrane-combine GGT and soluble GGT were significantly higher (P0.05) in hepatomas than those in adjacent paracancerous or and that distal cancerous tissues; and the frequencies of amplified fragment and hypomethylated M3 site of GGT -NC region genes were 100% and 75% in hepatomas, 85% and 55% in adjacent cancerous, and 75% and 50% in distal cancerous tissues, respectively. Conclusions The present data suggest that the abnormal expression of GGT proteins in hepatomas was related to hypomethylation status of GGT genes, and that the fragment analysis of the GGT genes might be a sensitive assay to monitor the hepatic cell canceration.Key words　Carci