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目的探讨EB病毒BARF1蛋白对胃癌细胞增殖与凋亡的影响。方法以空载体转染细胞SGC-pSG5为对照,采用细胞计数法和流式细胞术检测BARF1表达胃癌细胞系SGC7901(SGC-BARF1)的增殖能力。采用细胞计数法和Hoechst染色法分析紫杉醇诱导后的细胞凋亡状况。以实时荧光定量PCR和免疫印迹法分析细胞中c-met和Bcl-2基因表达的变化。结果与对照相比,BARF1表达细胞的增殖速度加快(P<0.05),处于细胞周期S期的细胞比例增高(P<0.01),抵抗紫杉醇诱导凋亡的能力明显增强(P<0.05),c-met和Bcl-2基因mRNA和蛋白质的表达量均显著增加,且二者呈正相关(P<0.01)。结论BARF1蛋白可能通过上调胃上皮细胞c-met和Bcl-2基因表达促进胃上皮细胞的增殖和抑制其凋亡,从而促进细胞的恶性转化。
Objective To investigate the effect of EBV BARF1 protein on proliferation and apoptosis of gastric cancer cells. Methods The empty vector transfected cells SGC-pSG5 as a control, the cell counting method and flow cytometry was used to detect the proliferation of gastric cancer cell line SGC7901 (SGC-BARF1). Cell apoptosis and Hoechst staining were used to analyze the apoptosis after paclitaxel induction. The changes of c-met and Bcl-2 gene expression in cells were analyzed by real-time fluorescence quantitative PCR and Western blotting. Results Compared with the control group, BARF1-expressing cells proliferated faster (P <0.05), the proportion of cells in S phase increased (P <0.01), and the ability of paclitaxel-induced apoptosis increased significantly The expressions of -met and Bcl-2 mRNA and protein were significantly increased (P <0.01). Conclusion BARF1 protein may promote the proliferation of gastric epithelial cells and inhibit the apoptosis of gastric epithelial cells by up-regulating the expression of c-met and Bcl-2 genes in gastric epithelial cells, thereby promoting malignant transformation of cells.