腺病毒介导p55γ基因N末端的过表达对胃癌细胞增殖和迁移的抑制  

Inhibitory Effects of p55γ Gene N-terminal Overexpression Mediated by Adenovirus on Proliferation and Migration of Gastric Carcinoma Cells

作　　者：冯丽[1] 孙晓杰[1] 高涵[1] 李珅[1] 张梅[1] 

机构地区：[1]齐齐哈尔医学院生化与分子生物学教研室,黑龙江齐齐哈尔161006

出　　处：《癌变．畸变．突变》2009年第6期409-412,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：黑龙江省自然科学基金资助项目(ZA2006-04);黑龙江省卫生厅科研项目(2006-418)

摘　　要：背景与目的:通过腺病毒介导,观察PI3K P55γ调节亚基N末端对胃癌细胞增殖和迁移的影响。材料与方法:在腺病毒E1基因转化的人胚肾细胞HEK293中扩增复制缺陷型重组腺病毒Ad-N24-GFP以及空载体病毒Ad-GFP,分别用Ad-GFP和Ad-N24-GFP感染MGC803细胞后,通过荧光显微镜观察病毒对肿瘤细胞的感染效率,计数活细胞,制作细胞生长曲线,观察Ad-N24-GFP对细胞生长的影响;并采用流式细胞术和细胞迁移实验分别检测Ad-N24-GFP对MGC803细胞周期和细胞运动迁移能力的影响。结果:重组腺病毒经过感染HEK293细胞后大量扩增,在病毒感染复数(MOI)为100时病毒对细胞的感染效率最高。与感染空载体的细胞相比,感染Ad-N24-GFP的MGC803细胞生长速度减慢、细胞倍增时间延长;Ad-N24-GFP的过表达使MGC803细胞G0/G1期百分率由(68.7±5)%上升到(84.2±5.4)%,差异具有统计学意义(P<0.05);感染Ad-N24-GFP的MGC803细胞其细胞迁移能力明显降低,与空载体组细胞相比,差异亦具有统计学意义(P<0.05)。结论:腺病毒介导p55γN末端24肽的过表达能够显著抑制胃癌MGC803细胞的增殖和迁移,其在胃癌的基因治疗上可能具有潜在的应用前景。BACKGROUND AND AIM： To investigate the effects of overexpression of the 24-amino-acid N-terminal end of p55γ regulatory subunit of phosphoinositide-3 kinase on proliferation and migration mediated by adenovirus in MGC803 gastric carcinoma cells. MATERIALS AND METHODS： The recombinant adenovirus-Ad-N24-GFP and control virus Ad-GFP were amplified in HEK293 cells. The virus infection rate of tumor cells was determined by fluorescence microscope. The effects of Ad-N24-GFP on cell proliferation and cell cycle were detected by cell growth curve and flow cytometry. The effect of Ad-N24-GFP on cell migration was detected by Transwell migration assay. RESULTS： The infection rate of recombinant adenovims of MGC803 cells was highest when MOI = 100. Compared with control cells, the growth of MGC803 cells after Ad-N24-GFP infection was suppressed and cell doubling time was prolonged. The percentage of Ad-N24-GFP cells in G0/G1 phase was （84.2 ±5.4） %, significantly higher than that of control cells in G0/G1 phase （68.7± 5）% （P 〈 0.05）. Overexpression of the Ad-N24-CFP fusion protein markedly resulted in decrease of MGC803 cell migration compared with the control cells （P 〈 0.05）. CONCLUSION： Overexpression of the p55γ gene N-terminal 24 peptide mediated by adenovirus inhibited cell proliferation and migration in gastric carcinoma MGC803 cells. It could have potential application in gastric carcinoma gene therapy.

关 键 词：腺病毒 p55γ 胃癌 细胞增殖 迁移 

分 类 号：R733[医药卫生—肿瘤]