Stability of HBV DNA in cell lines and nude mouse-passaged tissues derived from human hepatocellular carcinoma.

M R Chen, T Y Hsu, M J Chou, A C Chang, J Y Chen, C S Yang
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Abstract

Human hepatitis B virus (HBV) infection has been closely linked to the occurrence of hepatocellular carcinoma (HCC). Hepatoma cell lines and nude mouse-passaged hepatoma tissues were used in this report to study the HBV DNA status in these cells after passage. DNA was extracted from seven hepatoma cell lines and three nude mouse passaged HCC lines. Southern blot hybridization technique was performed with either cloned HBV whole genome or subgenomic DNA fragments as probes to analyze the presence of HBV DNA. Integration of HBV DNA fragments was detected in one mouse passaged tissue, R. Hybridization with HBV subgenomic DNA revealed that there were some DNA rearrangements of the integrated HBV DNA in R. However, the integrated HBV DNA could not be detected in the cell line derived from R after in vitro cultivation for 2 years. Both episomal form and integrated HBV DNA were detected in a cell line NTU-h3. Episomal form HBV DNA ih NTU-h3 changed after several passages. HBV DNA in NTU-h3 was unstable after in vitro cultivation. Therefore, we concluded that the presence of HBV DNA might not be essential for the maintenance of the tumorigenicity of hepatoma and the nude mouse system was more stable for maintaining HBV DNA in HCC.

人肝细胞癌细胞系和裸鼠传代组织中HBV DNA的稳定性。
人类乙型肝炎病毒(HBV)感染与肝细胞癌(HCC)的发生密切相关。本报告利用肝癌细胞系和裸鼠传代肝癌组织,研究了这些细胞传代后HBV DNA的状态。从7株肝癌细胞株和3株裸鼠传代肝癌细胞株中提取DNA。用克隆的HBV全基因组或亚基因组DNA片段作为探针,采用Southern blot杂交技术分析HBV DNA的存在。在一个小鼠传代组织R中检测到HBV DNA整合片段,与HBV亚基因组DNA杂交发现R中整合的HBV DNA存在一些DNA重排,但在体外培养2年后,在R衍生的细胞株中未检测到整合的HBV DNA。在细胞系NTU-h3中检测到外体形式和整合HBV DNA。带NTU-h3的Episomal形式HBV DNA在几次传代后发生了变化。NTU-h3体外培养后HBV DNA不稳定。因此,我们得出结论,HBV DNA的存在可能不是维持肝癌致瘤性所必需的,在HCC中,裸鼠系统对于维持HBV DNA更为稳定。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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