摘要
目的:探讨巨噬细胞加帽蛋白(macrophage-capping protein,CapG)对胃癌细胞系迁移和增殖能力的影响。方法:采用real-time PCR方法检测了4种胃癌细胞AGS、BCG823、PHM82、MNK45中CapG基因的表达情况,选择低表达并且易转染的AGS细胞作为研究对象,设计针对CapG的特异性引物并合成重组质粒。构建可以表达CapG的慢病毒包装系统,通过侵染人胃癌细胞系AGS,建立可稳定表达CapG的细胞株。通过CCK8实验分析过表达CapG基因对AGS细胞的生长和增殖能力的影响,细胞划痕以及Transwell小室实验分析过表达CapG基因对AGS细胞迁移能力的影响。结果:过表达CapG后,AGS细胞的生长速度略低于对照组,但二者间差异无统计学意义(t=2.424,P=0.073)。划痕试验显示CapG实验组划痕间距相对于对照组明显缩小,两组平均缩小距离分别为336.99μm和45.54μm,差异有统计学意义(t=14.97,P=0.004)。Transwell试验显示CapG实验组和对照组穿膜细胞数目分别为176个和70个,CapG实验组显著多于对照组,差异有统计学意义(t=40.00,P<0.001)。结论:过表达CapG基因对胃癌细胞系AGS细胞的生长和增殖无显著影响,过表达CapG基因能促进胃癌细胞系AGS细胞的迁移。
Objective: To investigate the effect of macrophage-capping protein( CapG) on migration and proliferation of human gastric cancer cell line. Methods: Real-time PCR method was used to detect the expression of CapG gene in four gastric cancer cell lines,and AGS cells with low expression and transfection were selected as the research objects. Specific primers were designed for CapG and recombinant plasmids synthesized. A lentivirus packaging system which could express CapG was constructed,and a cell line stably expressing CapG was established by infecting human gastric cancer cell line AGS cells.The effect of overexpression of CapG gene on the growth and proliferation of AGS cells was analyzed by CCK8 assay. Cells cratch and Transwell assay were used to analyze the effect of overexpression of CapG gene on AGS cell migration. Results: After the overexpression of CapG,the growth rate of AGS cells was slightly lower than that of the control group,but there was no significant difference between the two groups( t = 2. 424,P = 0. 073). Scratch test showed that the average narrowing distance of the scratches in the CapG experimental group was significantly reduced compared with the control group,the average narrowing distance of the CapG experimental group and the control group was 336. 99 μm and 45. 54 μm,the difference was statistically significant( t = 14. 97,P = 0. 004). The average number of cell penetrating membrane in the CapG experimental group and the e GFP control group was 176 and 70,the number of the cells in the CapG experimental group was significantly higher than that of the control group( t =40. 00,P < 0. 001). Conclusion: The overexpression of CapG gene has no significant effect on the growth and proliferation of AGS cells of gastric cancer cell line. Overexpression of CapG gene can promote the migration of AGS cells of gastric cancer cell lines.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2017年第3期489-494,共6页
Journal of Peking University:Health Sciences
基金
国家自然科学基金(81450024)资助~~
关键词
巨噬细胞加帽蛋白
胃肿瘤
细胞系
肿瘤
细胞增殖
细胞运动
Macrophage-capping protein
Stomach neoplasms
Cell line,tumor
Cell proliferation
Cell movement
