Wu Jing, Qin Yang, Li Bo, He Wen-zhi, Sun Zhi-lin
Department of Biochemistry and Molecular Biology, School of Preclinical and Forensic Medicine, West China Medical Center, Chengdu 610041, Sichuan Province, China.
Genomics. 2008 May;91(5):443-50. doi: 10.1016/j.ygeno.2008.01.007. Epub 2008 Mar 20.
In this study, 39 human hepatocellular carcinoma (HCC) tissues and 7 normal adult liver tissues were screened for heterozygous polymorphisms in IGF2, H19, and the differentially methylated region of H19 (H19DMR) using PCR-RFLP and PCR sequencing. The imprinting of IGF2 and H19 was examined by RT-PCR-RFLP, while the methylation profile of H19DMR was detected by bisulfite sequencing from every informative sample. Of the informative HCC samples 47.06% (8 of 17) demonstrated a gain of imprinting of IGF2, and 21.74% (5 of 23) of the informative HCC samples demonstrated a loss of imprinting of H19. Interestingly, we found three methylation profiles for H19DMR in the informative HCC samples: hyper-, medium-, and hypomethylated profiles. Furthermore, the hypomethylated and hypermethylated profiles were immediately associated with aberrant imprinting of IGF2 and H19.
在本研究中，使用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）和PCR测序技术，对39例人类肝细胞癌（HCC）组织和7例正常成人肝脏组织进行了胰岛素样生长因子2（IGF2）、母系表达基因1（H19）及H19差异甲基化区域（H19DMR）杂合多态性的筛查。采用逆转录PCR-限制性片段长度多态性（RT-PCR-RFLP）检测IGF2和H19的印记，同时通过亚硫酸氢盐测序检测每个信息样本中H19DMR的甲基化谱。在有信息的HCC样本中，47.06%（17例中的8例）表现出IGF2印记增强，21.74%（23例中的5例）有信息的HCC样本表现出H19印记缺失。有趣的是，我们在有信息的HCC样本中发现了H19DMR的三种甲基化谱：高甲基化、中等甲基化和低甲基化谱。此外，低甲基化和高甲基化谱与IGF2和H19的异常印记立即相关。
