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目的建立稳定、高效的HBx蛋白体外表达细胞株,以便进一步研究HBX基因和HBx蛋白的致癌作用及机理。方法建立HBX真核表达载体pcDNA3.1-HBX,通过脂质体介导将pcDNA3.1-HBX转染到人正常肝细胞株HL7702及人肝肿瘤细胞株HepG2中。结果 RT-PCR、蛋白质印迹及免疫荧光的实验结果显示,HBX基因在人正常肝细胞株HL7702及人肝肿瘤细胞株HepG2中得到了表达。结论成功构建了表达HBx蛋白的HL7702-HBX与HepG2-HBX细胞株,为进一步研究HBX基因和HBx蛋白的致癌作用及机制奠定了实验基础。
Objective To establish a stable and efficient cell line expressing HBx protein in vitro so as to further study the carcinogenic effects and mechanisms of HBx gene and HBx protein. Methods HBX eukaryotic expression vector pcDNA3.1-HBX was constructed and transfected into human normal liver cell line HL7702 and human hepatoma cell line HepG2 by lipofectamine. Results The results of RT-PCR, Western blotting and immunofluorescence showed that HBX gene was expressed in human normal liver cell line HL7702 and human hepatoma cell line HepG2. Conclusion The HL7702-HBX and HepG2-HBX cell lines expressing HBx protein were successfully constructed, which laid the foundation of further study on carcinogenesis and mechanism of HBx gene and HBx protein.