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细胞外信号调节激酶(ERK)级联是丝裂原活化蛋白激酶(MAPK)通路中的重要一员,与细胞增殖、分化、凋亡关系密切。乙型肝炎病毒(HBV)x(HBx)基因是一种致肝癌的重要因子,能激活肝细胞内ERK通路,从而影响细胞生物学特性。本实验通过稳定表达HBx的HepG2细胞(HBx+HepG2)对顺铂以及顺铂联合MAPK/ERK激酶(MEK)特异性抑制剂PD98059敏感性差异的研究,探讨PD98059提高HBx+HepG2 对顺铂敏感性的作用机制。一、材料与方法1.质粒、细菌及细胞株:质粒pCP10由北京解放军第三0二医院传染病研究所成军教授惠赠,内含两拷贝的HBV ayw 亚型全长基因组序列;pcDNA3真核表达载体购于美国
Extracellular signal-regulated kinase (ERK) cascade is an important member of mitogen-activated protein kinase (MAPK) pathway, which is closely related to cell proliferation, differentiation and apoptosis. Hepatitis B virus (HBV) x (HBx) gene is an important factor in hepatocellular carcinoma, which can activate the intracellular ERK pathway in hepatocytes and thus affect the biological characteristics of cells. In this study, HBx + HepG2 cells stably expressing HBx (HepG2) on cisplatin and cisplatin combined with MAPK / ERK kinase (MEK) specific inhibitor PD98059 sensitivity to explore the PD98059 increase HBx + HepG2 sensitivity to cisplatin The mechanism of action. Materials and Methods 1. Plasmids, Bacteria and Cell Lines: Plasmid pCP10 was kindly donated by Professor Cheng Jun from the Institute of Infectious Diseases of the Third Hospital of PLA of Beijing, containing two copies of the full-length genomic sequence of HBV ayw subtype. The pcDNA3 eukaryotic The expression vector was purchased from the United States