肝癌细胞系中SLIT2、DAPK、RIZ1、CHFR、EDNRB、3-OST-2基因甲基化状态

Methylation of SLIT2,DAPK,RIZ1,CHFR,EDNRB,3-OST-2 genes in hepatocellular carcinoma cell lines

目的了解SUN449、BEL-7402、SMMC-7721、Hep3B和HepG2 5种肝癌细胞系中SLIT2、DAPK、RIZ1、CHFR、ED-NRB和3-OST-2 6种基因启动子区域甲基化状态。方法体外培养以上5种肝癌细胞系,应用甲基化特异性PCR(MSP)技术,检测5种肝癌细胞系中SLIT2、DAPK、RIZ1、CHFR、EDNRB和3-OST-2 6种基因启动子区域的甲基化状态。结果 SLIT2、RIZ1、EDNRB和3-OST-2 4种基因启动子区域在5种肝癌细胞系SUN449、BEL-7402、SMMC-7721、Hep3B和HepG2中为完全甲基化,甲基化率均为100%;DAPK基因在此5种肝癌细胞系中为完全非甲基化,未发现甲基化状态,甲基化率为0;CHFR基因启动子区在SUN449、BEL-7402、SMMC-7721和HepG2细胞系中发生甲基化,甲基化率为80%,在SUN449、SMMC-7721和HepG2中为不完全甲基化,在BEL-7402细胞系中为完全甲基化,在Hep3B中为完全非甲基化。结论肝癌细胞系中SLIT2、DAPK、RIZ1、CHFR、EDNRB和3-OST-2基因启动子区的高甲基化是普遍现象,而DAPK基因呈低甲基化水平。

Objective To study the methylation of SLIT2DAPK,RIZ1,CHFR,EDNRB and 3-OST-2 genes in 5 hepatocellular carcinoma（HCC） cell lines-SUN449,BEL-7402,SMMC-7721,Hep3B and HepG2.Methods Methylation-specific PCR was adopted to detected the promoter methylation of these genes in the 5 HCC cell lines cultured in vitro.Results SLIT2,RIZ1,EDNRB and 3-OST-2 were completely showing methylation in SUN449,BEL-7402,SMMC-7721,Hep3B and HepG2.The methylation rate was 100%;DAPK was completely unmethylation,and the methylation rate was 0;Methylation of CHFR was deteceted in SUN449,BEL-7402,SMMC-7721 and HepG2,The methylation rates were 80%,Methylation was deteceted partly in SUN449,SMMC-772 and HepG2,but completely methylation in BEL-7402 and completely unmethylation in Hep3B.Conclusion Methylation of SLIT2,DAPK,RIZ1,CHFR,EDNRB and 3-OST-2 is a common event in 5 HCC cell lines,but the methylation rate of DAPK is low.