热休克蛋白gp96表达对B16F10细胞生长的影响

Effect of Heat Shock Protein gp96 on the Growth of B16F10 Cells

目的 探讨热休克蛋白gp96表达对B16F10细胞株生物学行为的影响。 方法 构建gp96正义和反义真核重组表达质粒 pcDNA hgp96和 pcDNA anti96P ,并转染B16F10细胞 ,G 418筛选稳定转染的阳性细胞克隆 ,然后Western印迹分析细胞gp96的表达情况 ,绘制细胞生长曲线 ,观察 gp96表达对细胞生长特性的影响。 结果 转染了 gp96正义表达质粒的B16F10细胞gp96表达水平升高 ,细胞生长缓慢 ,倍增时间延迟 ;反义gp96表达质粒明显抑制了B16F10细胞 gp96表达 ,使细胞倍增时间提前。结论 热休克蛋白 gp96表达水平升高可抑制肿瘤细胞的生长 ,而 gp96表达水平下降则肿瘤细胞生长速度加快 。

Objective To investigate the effect of heat shock protein gp96 on the biological behavior of cancer cells.Methods The sense and anti-sense recombinant plasmids pcDNA-hgp96 and pcDNA-anti96P were constructed by ligating the fragment of gp96 cDNA into pcDNA3,a eukaryotic expressing vector.Then the plasmids were transfected into B16F10 cells and the stable positive transfectants were selected with G418.The expression of gp96 in cells was detected by Western blotting.The cell growth curves were made to investigate the effect of gp96 on the growth characteristics of B16F10 cells.Results The transfection of sense gp96 plasmid increased the expression of gp96 protein in B16F10 cells,slowed down the growth rate of the cells,and retarded the time that the cells doubly proliferated.The anti-sense gp96 recombinant inhibited significantly the expression of gp96 in B16F10 cells,and advanced the time that the cells doubly proliferated.Conclusion The increase of gp96 expression can inhibit the growth of cancer cells,and the decline of its expression can accelerate the growth of cancer cells.These indicate that gp96 may play an important role in tumorigenesis and development of cancer.