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[目的]探讨HBx突变体在肝细胞肝癌发病机制中的作用。[方法]收集慢性HBV感染者肝穿组织标本和HBV阳性的肝细胞肝癌组织标本,提取DNA。通过3D-PCR方法扩增HBx基因并测序检测HBx基因的突变。分析两种组织标本内HBx基因的突变模式,以及突变对HBx蛋白序列的影响。进而构建所检测到的HBx突变体的真核表达质粒,体外研究其生物学活性。[结果]在所检测的7例慢性HBV感染者肝穿标本内,有3例标本内检测到因G到A突变而产生的C端截短型的HBx突变体。在所检测的2例HBV阳性的肝细胞肝癌组织内,也检测到相似的C端截短型HBx突变体。体外研究这种C端截短型HBx突变体的生物学功能发现其具有促进细胞增殖的功能。[结论]C端截短型HBx突变体在HBV相关的肝细胞肝癌发病机制中具有重要意义。
[Objective] To investigate the role of HBx mutant in the pathogenesis of hepatocellular carcinoma. [Method] Chronic HBV infected liver tissue and HBV positive hepatocellular carcinoma tissue samples were collected to extract DNA. The HBx gene was amplified by 3D-PCR and sequenced to detect mutations in the HBx gene. The HBx gene mutation pattern in both tissue samples was analyzed and the effect of the mutation on the HBx protein sequence was analyzed. Furthermore, the eukaryotic expression plasmid of the detected HBx mutant was constructed and its biological activity was studied in vitro. [Results] The HBx mutants with C-terminal truncation due to G to A mutation were detected in 3 specimens of liver biopsy specimens from 7 chronic HBV infected patients. Similar C-terminal truncated HBx mutants were also detected in the two HBV-positive hepatocellular carcinoma tissues detected. In vitro studies of the biological function of this C-terminal truncated HBx mutant were found to have the function of promoting cell proliferation. [Conclusion] C-terminal truncated HBx mutant is of great significance in the pathogenesis of hepatocellular carcinoma associated with HBV.