HL7702-HBX与HepG2-HBX细胞株的建立及鉴定

Establishment and identification of in vitro HL7702 and HepG2 cell ines with HBX gene expresion

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。

Aim To establish in vitro cell line with stable and effective HBxAg expression,for study of carcinogenesis mechanism of HBX gene and HBxAg. Methods A recombinant plasmid （pcDNA3.1-HBX）containing the full length of HBX gene was transfected into HL7702 cells and HepG2 cells. The expression of HBx in the cells was analyzed by Western blot and immunofiuorescency using anti-HBx antibodies. Results Cell clones were obtained after being transfected with pcDNA3.1-HBX plasmid. Stable expression of HBxAg was detected by Western blot,immunofluorescency and RT-PCR. Conclusion HBX gene expression cell lines were constructed in this study. The property of this system ensures stable and high level expression of HBx. It is a useful tool for functional studies of HBX gene.