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目的:通过腺病毒介导,研究磷脂酰肌醇3-激酶p55γ调节亚基N末端24个氨基酸的过表达对胃癌MGC803细胞生长增殖的影响。方法:在HEK293细胞中扩增重组腺病毒Ad-N24-GFP以及空载体病毒Ad-GFP,并进行病毒滴度以及腺病毒对胃癌细胞感染率的测定,免疫印迹方法鉴定重组腺病毒感染MGC803细胞后融合蛋白N24-GFP的表达。通过细胞生长曲线和克隆形成实验观察Ad-N24-GFP对细胞增殖的影响。结果:重组腺病毒经过感染HEK293细胞后大量扩增,病毒滴度测定为1.0×1010pfu/ml,重组腺病毒对MGC803细胞的感染率在感染复数(MOI)为100时最强。与空载体细胞相比较,N24-GFP的高表达明显抑制MGC803细胞的生长,且抑制程度随病毒感染复数的增加呈逐渐增强的趋势;感染Ad-N24-GFP组细胞的克隆形成率[(28.30±3.25)%]明显低于对照组细胞[(42.16±2.84)%,P<0.05]。结论:腺病毒介导N24p55γ基因的过表达能抑制胃癌细胞MGC803的增殖,其在胃癌基因治疗上可能具有潜在的应用前景。
OBJECTIVE: To study the effect of 24 amino acids of N-terminal 24 amino acids of phosphatidylinositol 3-kinase (P55γ) regulatory subunit on the proliferation and proliferation of gastric cancer MGC803 cells by adenovirus mediated. Methods: The recombinant adenovirus Ad-N24-GFP and empty vector Ad-GFP were amplified in HEK293 cells and the titer of virus and the infection rate of adenovirus were detected. The recombinant adenovirus was used to infect MGC803 cells Post-fusion protein N24-GFP expression. The effect of Ad-N24-GFP on cell proliferation was observed by cell growth curve and clonogenic assay. Results: The recombinant adenovirus was amplified in HEK293 cells. The titer of the recombinant adenovirus was 1.0 × 1010 pfu / ml. The infection rate of recombinant adenovirus to MGC803 cells was the strongest at the MOI of 100. Compared with empty vector cells, the high expression of N24-GFP significantly inhibited the growth of MGC803 cells, and the degree of inhibition tended to increase with the increase of viral infections. The rate of clone formation in Ad-N24-GFP infected cells [(28.30 ± 3.25)%] was significantly lower than that in the control group [(42.16 ± 2.84)%, P <0.05]. Conclusion: Adenovirus-mediated N24p55γ gene overexpression can inhibit the proliferation of gastric cancer cell line MGC803, which may have potential applications in the gene therapy of gastric cancer.