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目的进一步研究ＨＢＶｘ基因产物ＨＢｘＡｇ与ｐ５３蛋白的相互关系，探讨其在原发性肝癌发生中的作用机制。方法以肝癌细胞株Ｈｅｐ３Ｂ为靶细胞，应用薄层层析法做报道基因氯霉素乙酰转移酶（ＣＡＴ）试验与ｐ５３和ＨＢｘ基因共转染，并构建了地塞米松诱导表达的ＨＢｘ基因质粒，用免疫荧光法观察ＨＢｘＡｇ表达对细胞内ｐ５３蛋白的影响。结果应用ＨＢｘ基因与ｐ５３和ｐＧ１３ＣＡＴ共转染，随ＨＢｘ量的增加而ＣＡＴ信号增强，免疫组化显示，当ＨＢｘＡｇ与ｐ５３蛋白在细胞内结合形成复合物后，ｐ５３蛋白进入胞核部分受阻滞，胞浆内ｐ５３蛋白含量增加。结论ＨＢｘＡｇ与ｐ５３存在蛋白质蛋白质结合的关系，使ｐ５３蛋白正常的负调节功能降低或失活，在ＨＢＶ感染相关的原发性肝癌发生中具有重要意义。
Objective To further study the relationship between HBVx gene product HBxAg and p53 protein and explore its mechanism of action in the development of primary liver cancer. Methods The hepatoma cell line Hep3B was used as the target cell. The chloramphenicol acetyltransferase (CAT) reporter gene was co-transfected with p53 and HBx gene by thin layer chromatography. The HBx gene plasmid induced by dexamethasone was constructed. The effect of HBxAg expression on intracellular p53 protein was observed by immunofluorescence. Results HBx gene was co-transfected with p53 and pG13CAT. The CAT signal was increased with the increase of HBx content. Immunohistochemistry showed that when HBxAg and p53 protein combine to form a complex in the cell, the p53 protein enters the nucleus partially blocked. The cytoplasmic p53 protein content increased. Conclusion There is a protein-protein binding relationship between HBxAg and p53, which makes the normal negative regulation function of p53 protein decrease or be inactivated. It plays an important role in the occurrence of primary liver cancer associated with HBV infection.