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目的探讨免疫负性调控基因A20在肝癌中的表达并构建A20真核表达载体。方法采用免疫组化技术,检测46例肝癌患者癌组织和癌旁组织中A20的表达情况,并进一步构建A20真核表达载体,测序正确后以脂质体法转染至SMMC-7721肝癌细胞系中,RT-PCR和Western blot检测转染后A20的mRNA和蛋白表达。结果免疫组化结果显示,A20在肝癌组织中的表达明显低于癌旁组织,差异具有统计学意义(P<0.05)。A20的表达与肝癌分级分化和肿瘤大小有关(P<0.05),成功构建A20真核表达载体,并在SMMC-7721肝癌细胞系中获得高表达。结论免疫负调控基因A20可能参与了肝癌的发生机制,成功构建的A20真核表达载体可作为进一步研究的重要工具。
Objective To investigate the expression of immunoregulatory gene A20 in hepatocellular carcinoma (HCC) and to construct A20 eukaryotic expression vector. Methods Immunohistochemistry was used to detect the expression of A20 protein in 46 cases of hepatocellular carcinoma and adjacent normal tissues. The A20 eukaryotic expression vector was further constructed and transfected into SMMC-7721 hepatocellular carcinoma cell line RT-PCR and Western blot were used to detect the mRNA and protein expression of A20 after transfection. Results The results of immunohistochemistry showed that the expression of A20 in hepatocellular carcinoma was significantly lower than that in paracancerous tissues, the difference was statistically significant (P <0.05). The expression of A20 was correlated with the differentiation and tumor size (P <0.05). The A20 eukaryotic expression vector was successfully constructed and was highly expressed in SMMC-7721 hepatoma cell line. Conclusions The immunoregulatory gene A20 may be involved in the pathogenesis of hepatocellular carcinoma. The successfully constructed A20 eukaryotic expression vector can be used as an important tool for further research.