论文部分内容阅读
目的探讨过表达高尔基体α-甘露糖苷酶Ⅱ(Golgiα-mannosidaseⅡ,GMⅡ)对人胃癌BGC-823细胞凋亡的影响。方法构建GMⅡ基因真核过表达质粒EX-E2372-M03,通过脂质体Lipofectamine 2000转染BGC-823细胞,用400 mg/LG418筛选稳定转染的细胞。采用RT-PCR和Western blot分别检测转染后细胞中GMⅡ基因mRNA的转录水平以及蛋白的表达水平,Hoechst 33258荧光染色法和流式细胞术(Annexin V/PI双染)检测过表达GMⅡ后细胞的凋亡情况。结果转染重组质粒EX-E2372-M03后,BGC-823细胞GMⅡ基因mRNA的转录水平和蛋白的表达水平明显增加(P<0.05);细胞凋亡率明显降低(P<0.05)。结论过表达GMⅡ可能通过抑制胃癌细胞的凋亡,进而促进胃癌的发生发展。
Objective To investigate the effect of Golgi α-mannosidase Ⅱ (GMⅡ) overexpression on the apoptosis of human gastric cancer BGC-823 cells. Methods The eukaryotic overexpression plasmid GM-E2372-M03 was constructed and transfected into BGC-823 cells by Lipofectamine 2000. Stably transfected cells were screened by 400 mg / L LG418. The transcript level and protein expression of GMⅡ gene were detected by RT-PCR and Western blot respectively. The expression of GMⅡ mRNA was detected by Hoechst 33258 staining and Annexin V / PI double staining. Apoptosis. Results Transfection of recombinant plasmid EX-E2372-M03 resulted in a significant increase in the mRNA and protein expression of GMⅡ mRNA in BGC-823 cells (P <0.05). The apoptotic rate was significantly decreased (P <0.05). Conclusion Overexpression of GMⅡ may promote the development of gastric cancer by inhibiting the apoptosis of gastric cancer cells.