Effect of hepatitis B virus X gene integration on expression of zinc finger protein ZBTB20 in chronic hepatitis B patients complicated with hepatocellular carcinoma

中华传染病杂志 Pub Date : 2019-09-15 DOI:10.3760/CMA.J.ISSN.1000-6680.2019.09.007

Zebao He, Qiuyue Chen, Jiansheng Zhu, Yang Lu, Hai-hong Zhao, Zheping Fang

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Abstract

Objective To investigate the effect of hepatitis B virus (HBV) X gene integration on expression of zinc finger protein ZBTB20 in chronic hepatitis B (CHB) patients complicated with hepatocellular carcinoma (HCC). Methods Eighteen CHB patients complicated with HCC who underwent surgical treatment in Taizhou Enze Medical Center Enze Hospital and Taizhou Central Hospital from July 2015 to June 2017 were enrolled. Samples of carcinoma tissue, para-carcinoma tissue and corresponding normal liver tissue were collected from each case. DNA was extracted from three kinds of tissue samples. HBV-Alu-polymerase chain reaction (PCR) was used to amplify the integrated HBVX fragments and their bilateral flanking sequences in human genomic DNA. The integrated HBV fragments were determined by PCR products sequencing. Protein was extracted from three kinds of tissue samples.The level of expression of ZBTB20 was detected by protein imprinting. Statistical analysis was performed using t test, analysis of variance, and χ2 test. Results Among the 18 CHB patients complicated with HCC, integration of HBVX gene was found in 13 carcinoma tissue samples, 16 para-carcinoma tissue samples and 9 corresponding normal liver tissue samples. The difference was statistically significant (χ2=6.353, P=0.037). Of the 18 patients, the protein expressions of ZBTB20 in carcinoma tissue, para-carcinoma tissue and corresponding normal tissue were (50.14±11.25)%, (40.71±7.17)% and (39.06±5.17)%, respectively, which was statistically different (F=9.420, P<0.01). HBVX gene integration was detected at ZBTB20 locus in five patients. The expression levels of ZBTB20 in patients with HBVX gene integration at this locus in carcinoma tissue, para-carcinoma tissue and corresponding, normal liver tissue were all significantly lower than those in patients without HBVX gene integration (carcinoma tissue (37.37±10.30)% vs (55.06±7.06)%, para-carcinoma tissue (32.06±2.61)% vs (44.04±5.24)%, corresponding normal tissue (34.66±5.59)% vs (40.76±4.04)%, t=4.205, 4.821 and 2.589, respectively, all P<0.05). Conclusions Incidence of HBVX integration in para-carcinoma tissue is higher than that in carcinoma tissue in CHB patients complicated with HCC.The expression level of ZBTB20 in carcinoma tissue is higher than that in para-carcinoma tissue. Integration of HBVX gene at ZBTB20 locus may decreases the expression of ZBTB20. Key words: Hepatitis B virus; Carcinoma, hepatocellular; Gene integration; Zinc finger protein ZBTB20

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乙型肝炎病毒X基因整合对慢性乙型肝炎合并肝癌患者锌指蛋白ZBTB20表达的影响

目的探讨乙型肝炎病毒(HBV) X基因整合对慢性乙型肝炎(CHB)合并肝细胞癌(HCC)患者锌指蛋白ZBTB20表达的影响。方法选取2015年7月至2017年6月在泰州市恩泽医疗中心恩泽医院和泰州市中心医院行手术治疗的18例CHB合并HCC患者。取癌组织、癌旁组织及相应的正常肝组织标本。从三种组织样本中提取DNA。采用hbv - alu聚合酶链式反应(PCR)扩增HBVX整合片段及其在人类基因组DNA中的双侧序列。聚合酶链反应产物测序检测HBV整合片段。从三种组织样本中提取蛋白质。采用蛋白印迹法检测ZBTB20的表达水平。采用t检验、方差分析和χ2检验进行统计学分析。结果在18例CHB合并HCC患者中，13例癌组织样本、16例癌旁组织样本和9例相应的正常肝组织样本中发现HBVX基因整合。差异有统计学意义(χ2=6.353, P=0.037)。18例患者癌组织、癌旁组织及相应正常组织中ZBTB20蛋白表达分别为(50.14±11.25)%、(40.71±7.17)%、(39.06±5.17)%，差异有统计学意义(F=9.420, P<0.01)。5例患者在ZBTB20位点检测到HBVX基因整合。在该位点整合HBVX基因的患者中，ZBTB20在癌组织、癌旁组织和相应的正常肝组织中的表达水平均显著低于未整合HBVX基因的患者(癌组织(37.37±10.30)% vs(55.06±7.06)%，癌旁组织(32.06±2.61)% vs(44.04±5.24)%，相应的正常组织(34.66±5.59)% vs(40.76±4.04)%，t分别=4.205、4.821和2.589,P均<0.05)。结论CHB合并HCC患者HBVX在癌旁组织的整合率高于癌组织。ZBTB20在癌组织中的表达水平高于癌旁组织。在ZBTB20位点整合HBVX基因可降低ZBTB20的表达。关键词:乙型肝炎病毒;癌,肝细胞;基因整合;锌指蛋白ZBTB20

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来源期刊

中华传染病杂志

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期刊介绍： The Chinese Journal of Infectious Diseases was founded in February 1983. It is an academic journal on infectious diseases supervised by the China Association for Science and Technology, sponsored by the Chinese Medical Association, and hosted by the Shanghai Medical Association. The journal targets infectious disease physicians as its main readers, taking into account physicians of other interdisciplinary disciplines, and timely reports on leading scientific research results and clinical diagnosis and treatment experience in the field of infectious diseases, as well as basic theoretical research that has a guiding role in the clinical practice of infectious diseases and is closely integrated with the actual clinical practice of infectious diseases. Columns include reviews (including editor-in-chief reviews), expert lectures, consensus and guidelines (including interpretations), monographs, short monographs, academic debates, epidemic news, international dynamics, case reports, reviews, lectures, meeting minutes, etc.