乙型肝炎患者肝细胞癌着丝粒蛋白K基因启动子区域甲基化水平的初步分析

doi:10.19871/j.cnki.xfcrbzz.2021.02.002

新发传染病电子杂志 ›› 2021, Vol. 6 ›› Issue (2): 84-87.doi: 10.19871/j.cnki.xfcrbzz.2021.02.002

乙型肝炎患者肝细胞癌着丝粒蛋白K基因启动子区域甲基化水平的初步分析

乐晓华¹, 刘琢冰³, 张香梅¹, 汪海燕³, 曾丽娇³, 郑娟³, 姜楠², 刘威龙³

1.深圳市第三人民医院病理科,广东深圳 518112;
2.深圳华侨城医院检验科,广东深圳 518020;
3.深圳市第三人民医院肝病研究所,广东深圳 518112

收稿日期:2020-12-29 出版日期:2021-05-31 发布日期:2021-06-24
通讯作者: 刘威龙,E-mail：weilong8380@hotmail.com
基金资助:
深科技创新项目（JCYJ20160427174431520）

Preliminary analysis of the methylation level of CENPK gene promoter region in hepatocellular carcinoma patients with hepatitis B

Le Xiaohua¹, Liu Zhuobing³, Zhang Xiangmei¹, Wang Haiyan³, Zeng Lijiao³, Zheng Juan³, Jiang Nan², Liu Weilong³

1. Department of Pathology, the Third People's Hospital of Shenzhen, Guangdong Shenzhen 518112, China;
2. Department of Laboratory Medicine, Shenzhen Overseas Chinese Town Hospital, Guangdong Shenzhen 518020, China;
3. Institute of Liver Diseases, the Third People's Hospital of Shenzhen, Guangdong Shenzhen 518112, China

Received:2020-12-29 Online:2021-05-31 Published:2021-06-24

摘要/Abstract

摘要： 目的比较乙型肝炎患者肝细胞癌及其癌旁肝组织中着丝粒蛋白K（CENPK）基因启动子区域甲基化水平的差异,提升肝癌早期诊断和治疗水平。方法从美国国家生物技术信息中心（NCBI）获取并分析CENPK基因的全长基因组序列,采用Primer 5.0软件设计针对CENPK基因启动子区域典型CpG岛的特异性扩增引物;利用亚硫酸氢盐测序聚合酶链反应（bisulfite sequencing PCR,BSP）分析比较4对肝细胞癌及其配对癌旁组织中CENPK基因启动子区域CpG岛的16个CG位点的甲基化情况。结果成功筛选及鉴定4对肝癌和癌旁组织中CENPK基因启动子区域CpG岛甲基化状态,经比较分析测序结果显示,3例乙型肝炎患者肝细胞癌组织中CENPK基因启动子区域CpG岛的甲基化水平显著低于癌旁组织（P<0.05）。结论 CENPK基因在肝细胞癌组织中异常高表达与其启动子区域的DNA低甲基化水平密切相关,提示表观遗传学机制在肝细胞癌发生发展中起到重要的作用。

关键词: 肝细胞癌, 着丝粒蛋白K, 甲基化, 乙型肝炎

Abstract: Objective To compare the methylation level of CENPK gene promoter region in HCC and its paracancerous liver tissue of patients with hepatitis B, and to improve the level of early diagnosis and treatment of liver cancer. Methods The full-length genome sequence of CENPK gene was obtained from NCBI, and specific primers for typical CpG islands in CENPK gene promoter region were designed using Primer 5.0 software. Bisulfite sequencing PCR(BSP) was used to analyze and compare the methylation of 16 CG sites in CpG island of CENPK promoter region in 4 pairs of HCC and its corresponding paracancerous tissues. Results The methylation status of CpG island in CENPK promoter region was successfully screened and identified in 4 pairs of hepatocellular carcinoma and paracancerous tissues. The results of comparative analysis and sequencing showed that the methylation level of CpG Island in CENPK promoter region in 75% (3/4) hepatitis B patients was significantly lower than that in paracancerous tissues (P<0.05). Conclusions The abnormal high expression of CENPK gene in hepatocellular carcinoma is closely related to the level of DNA hypomethylation in the promoter region, suggesting that epigenetic mechanisms play an important role in the development of hepatocellular carcinoma.

Key words: Hepatocellular carcinoma, Centromere protein K, Methylation, High expression, Hepatitis B

乐晓华, 刘琢冰, 张香梅, 汪海燕, 曾丽娇, 郑娟, 姜楠, 刘威龙. 乙型肝炎患者肝细胞癌着丝粒蛋白K基因启动子区域甲基化水平的初步分析[J]. 新发传染病电子杂志, 2021, 6(2): 84-87.

Le Xiaohua, Liu Zhuobing, Zhang Xiangmei, Wang Haiyan, Zeng Lijiao, Zheng Juan, Jiang Nan, Liu Weilong. Preliminary analysis of the methylation level of CENPK gene promoter region in hepatocellular carcinoma patients with hepatitis B[J]. Electronic Journal of Emerging Infectious Diseases, 2021, 6(2): 84-87.

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乙型肝炎患者肝细胞癌着丝粒蛋白K基因启动子区域甲基化水平的初步分析

Preliminary analysis of the methylation level of CENPK gene promoter region in hepatocellular carcinoma patients with hepatitis B

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