Wang Jing, Lin Jusheng, Chang Ying, Li Peiyuan, Yang Yuzhen
Institute of Liver Diseases, Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Aug;30(4):425-9. doi: 10.1007/s11596-010-0443-3. Epub 2010 Aug 17.
A novel HBV integration site involved in hepatocarcinogenesis was investigated. The HBV DNA integration sites were detected by Alu-PCR in hepatocellular carcinoma tissues, matched surrounding liver tissues in 30 patients with hepatitis B-related hepatocellular carcinoma (HCC) and 3 cases of normal liver tissues. The integration sites and flanking sequences in human genome were sequenced and blasted, and the expression of integrated HBV genes was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The influence of the up-regulated expression of integrated genes on hepatocarcinogenesis was analyzed. Nineteen integration sites of HBV DNA into HCC tissues were obtained by RT-PCR and sequencing. These genes encoding proteins were: LOC51030, LOC157777, minichromosome maintenance complex component 3 associated protein (MCM3AP), MCTP1, SH3 and multiple ankyrin repeat domains 2 isoform 2, CCDC40, similar to HCG2033532, mitochondrial ribosomal S5 pseudogene 4. One of them was integrated into the intron of MCM3AP. RT-PCR demonstrated that the expression levels of MCM3AP mRNA in HCC tissues, matched surrounding liver tissues and normal liver tissues were in a descendent order. The ratio of MCM3AP mRNA to the GAPDH mRNA in these three tissues was 1.07375, 0.21573, 0.06747 respectively, with the difference being statistically significant among them (P<0.05). Meanwhile, the expression levels of MCM3AP mRNA from HCC tissues in which HBV DNA integrated into MCM3AP were still significantly higher than those without HBV DNA integrated into MCM3AP. It was concluded that the HBV DNA integration sites into human genome were random, and MCM3AP was a new site. The up-regulated MCM3AP mRNA may affect flanking sequences which promote the hepatocarcinogenesis.
对一个参与肝癌发生的新型乙肝病毒（HBV）整合位点进行了研究。采用Alu-PCR技术在30例乙型肝炎相关肝细胞癌（HCC）患者的肝癌组织、配对的周围肝组织以及3例正常肝组织中检测HBV DNA整合位点。对人类基因组中的整合位点及其侧翼序列进行测序和比对，并通过逆转录聚合酶链反应（RT-PCR）检测整合的HBV基因的表达。分析整合基因上调表达对肝癌发生的影响。通过RT-PCR和测序获得了19个HBV DNA在肝癌组织中的整合位点。这些编码蛋白质的基因有：LOC51030、LOC157777、微小染色体维持复合体组分3相关蛋白（MCM3AP）、MCTP1、SH3和多个锚蛋白重复结构域2亚型2、CCDC40、类似于HCG2033532、线粒体核糖体S5假基因4。其中一个整合到了MCM3AP的内含子中。RT-PCR结果显示，MCM3AP mRNA在肝癌组织、配对的周围肝组织和正常肝组织中的表达水平呈下降趋势。这三种组织中MCM3AP mRNA与GAPDH mRNA的比值分别为1.07375、0.21573、0.06747，三者之间差异具有统计学意义（P<0.05）。同时，HBV DNA整合到MCM3AP的肝癌组织中MCM3AP mRNA的表达水平仍显著高于未整合HBV DNA的肝癌组织。结论是，HBV DNA整合到人类基因组中的位点是随机的，MCM3AP是一个新位点。上调的MCM3AP mRNA可能影响侧翼序列，从而促进肝癌的发生。
