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目的:研究乙型肝炎病毒X蛋白(HBx)羧基端31个氨基酸自然缺失突变体(HBxΔ31)对肝癌细胞蛋白质表达的影响。方法:脂质体法介导全长HBx基因和HBxΔ31突变体表达载体转染人肝癌细胞系Hep G2,Western blotting分析证实HBx蛋白的表达,双向电泳分离细胞质蛋白质,图像分析Hep G2-HBx及Hep G2-HBxΔ31的差异蛋白表达,MALDI-TOF-MS及数据库搜索鉴定差异蛋白质。结果:在Hep G2-HBxΔ31及Hep G2-HBxΔ31细胞株中筛选鉴定出6个差异表达蛋白质,分别为HSPB1、Ran BP1、Rho-GDI alpha、Serpin B5、Cap Z beta、MRP-L12,在HBxΔ31-Hep G2细胞株中MRPL12、Ran BP1、HSPB1高表达,RhoGDI alpha、Serpin B5、Cap Z beta低表达。结论:HBx蛋白羧基端31个氨基酸突变缺失会改变肝癌细胞的蛋白质表达,这些蛋白与肿瘤的形成及转移密切相关,或可成为肝癌诊断及预后的标志物。
Objective: To study the effect of HBxΔ31, a 31 amino acid carboxyl terminal deletion of hepatitis B virus X protein (HBxΔ31), on the protein expression of hepatocellular carcinoma cells. Methods: The full-length HBx gene and HBxΔ31 mutant vector were transfected into human hepatocellular carcinoma cell line Hep G2 by liposome method. The expression of HBx protein was confirmed by Western blotting, and the cytoplasmic proteins were separated by two-dimensional electrophoresis. Hep G2-HBx and Hep Differential protein expression of G2-HBxΔ31, MALDI-TOF-MS and database search identified differential proteins. RESULTS: Six differentially expressed proteins were identified in Hep G2-HBxΔ31 and Hep G2-HBxΔ31 cell lines, including HSPB1, Ran BP1, Rho-GDI alpha, Serpin B5, Cap Z beta and MRP-L12, Hep G2 cell lines MRPL12, Ran BP1, HSPB1 high expression, RhoGDI alpha, Serpin B5, Cap Z beta low expression. Conclusion: The deletion of the 31 amino acid mutation in the carboxyl end of HBx protein will change the protein expression of hepatocellular carcinoma cells. These proteins are closely related to tumor formation and metastasis, or may be the markers of diagnosis and prognosis of liver cancer.