肝细胞肝癌谷胱甘肽硫转移酶P1基因型和表型及其活性研究  

Study on the polymorphisms and promoter methylation and expression of the glutathione S-transferases P1 gene in hepatocellular carcinoma

作　　者：张友才[1] 陈永平[1] 陈金霞[1] 董培红[1] 邓长生[2] 

机构地区：[1]温州医学院附属第一医院感染内科,325005 [2]武汉大学中南医院消化内科

出　　处：《中华消化杂志》2006年第7期468-472,共5页Chinese Journal of Digestion

基　　金：湖北省科技攻关计划课题资助项目(2002AA301C84)

摘　　要：目的 探讨谷胱甘肽硫转移酶（GST）P1活性及基因多态、启动子甲基化与肝细胞肝癌（HCC）的关系.方法 紫外分光光度法检测HCC肿瘤细胞和癌旁肝细胞胞质中GSTP1活性,用甲基化特异性PCR技术检测GSTP1甲基化,PCR-RFLP技术检测53例HCC患者和74例健康人外周血基因组GSTP1基因多态性.结果 GSTP1三种基因型频率在病例组与对照组间差异无统计学意义（χ^2=0.84,v=2,P=0.656）.GSTP1甲基化率在肿瘤组织与癌旁组织间差异有统计学意义（χ^2=19.08,P＜0.01）,在Ⅲ～Ⅳ期肿瘤中的频率显著高于Ⅰ～Ⅱ期肿瘤（χ^2=4.84,P=0.028）.HCC肿瘤细胞胞质中GSTP1活性显著低于癌旁肝细胞（t=2.49,P=0.014）,Ⅰ～Ⅱ期肿瘤胞质中GSTP1活性显著高于Ⅲ～Ⅳ期肿瘤（t=2.31,P=0.025）,GSTP1甲基化的肿瘤细胞胞质中GSTP1活性显著低于未甲基化的肿瘤细胞（t=3.50,P=0.001）.结论 由GSTP1基因甲基化引起的GSTP1活性下降可能与肝细胞的癌性转变有关.Objective To study the relationships between hepatocellular carcinoma （HCC） and the polymorphisms, promoter methylation, and expression of glutathione S-transferases P1 gene （GST） P1 gene. Methods Using methylation-special PCR （MSP）, the methylated status of CpG islands of GSTP1 gene in tumor tissues of 53 HCC and its adjacent nontumor tissues were studied. The enzyme activities of GSTP1 were evaluated by ultraviolet colormetry. And using PCR-RFLP, the genetic polymorphisms of the GSTP1 genes of 74 healthy controls and 53 HCC patients were studied. Results The diffe-rences of the frequency of GSTP1 Ile/Ile, Ile/Val and Val/Val genotypes between HCC patients and the normal controls did not reach statistical significance （X^2 = 0.84, v = 2, P = 0. 656）. The frequency of methylation of CpG islands of GSTP1 gene was significantly higher among the HCC tumor tissues when compared to the corresponding nontumor tissues （X^2 = 19.08, P 〈 0. 001）, and significantly higher in stage Ⅲ-Ⅳ cases when compared to the stage Ⅰ -Ⅱ cases （X^2 = 4.84, P = 0. 028）. GSTP1 enzyme activities of cytoplasm in tumor cells were lower significantly than that in the adjacent nontumor tissues （t =2.49, P = 0. 014）, and significantly higher in stage Ⅰ- Ⅱ cases when compared to the stage Ⅲ-Ⅳ eases （t = 2.31, P= 0. 025）. On the other hand, the GSTP1 enzyme activities of cytoplasm in tumor cells with methylated status of GSTP1 gene were significantly lower than that in tumor cells with unmethylation （t = 3.50, P = 0. 001）. Conclusion GSTP1 inactivation via CpG island hypermethylation may contribute to the pathogenesis of HCC.

关 键 词：肝细胞肝癌 谷胱甘肽硫转移酶P1 多态性 甲基化 

分 类 号：R735.7[医药卫生—肿瘤] R5[医药卫生—临床医学]