摘要
目的明确Abl相互作用蛋白1（ABl1）在正常胃黏膜细胞系GES-1和胃癌细胞系AGS中的表达模式，并探讨ABl1过表达对AGS细胞体外增殖的影响。方法首先用免疫组化、免疫荧光、逆转录（RT）-PCR、实时定量PCR及Western印迹法明确ABl1在GES-1及AGS细胞中的表达模式；其次用脂质体转染方法分别将基因重组表达载体鼠干细胞病毒（MSCV）．绿色荧光蛋白（GFP）-ABl1质粒和对照载体MSCV-GFP质粒导人AGS细胞，用嘌呤霉素筛选稳定表达MSCV—GFP—ABl1和MSCV·GFP的细胞系，用实时定量PCR和Western印迹法鉴定是否成功构建了ABl1过表达的细胞模型；最后用细胞活力检测试剂盒[八肽胆囊收缩素（CCK-8）]检测ABII过表达对细胞增殖的影响。结果ABll在正常胃黏膜细胞系GES．1及胃癌细胞系AGS中均有表达；与GES．1细胞相比，ABll在AGS细胞中的表达从mRNA水平到蛋白质水平均下调；成功构建稳定表达MSCV．GFP．ABl1的AGS．MSCV—GFP—ABl1细胞模型，AGS—MSCV—GFP—ABll细胞中ABll的表达无论mRNA水平还是蛋白质水平均明显高于AGS细胞（均P=0．002）；CCK．8法检测显示，AGS．MSCV．GFP．ABl1细胞在24、48、72、96h4个时间点增殖率分别为1．46±0．31、4．75±0．12、6．62±0．32、8．96±0．27，均明显低于AGS细胞及AGS—MSCV—GFP细胞（均P〈0．01）。结论ABll在胃癌细胞中表达下调，其过表达可以抑制人胃癌细胞系AGS的增殖，提示ABll表达下调可能参与了胃癌的发生发展过程，而影响胃癌细胞的增殖可能是其发挥作用的重要途径之一。
Objective To investigate the expression of Abl-interacting protein 1 （ABI1） in normal gastric mucosal cell line GES-1 and gastric cancer cell line AGS, and the effects of ABI1 gene overexpression upon the proliferation of human gastric cancer cell AGS in vitro. Methods Firstly the ABI1 expression in GES-1 and AGS cells were identified by immunohistochemistry, immunofluorescene, RT-PCR, real-time PCR and Western blot. Secondly human gastric cancer cell line AGS was cultured and transfected with recombinant MSCV-GFP-ABI1 plasmid or blank plasmid MSCV-GPF. Real-time PCR and Western blot were used to detect the mRNA and protein expression of ABI1. And lastly the cell proliferation was detected by CCK-8 assay. Results ABI1 was expressed both in normal gastric mucosal cell line GES-1 and in gastric cancer cell line AGS. Compared to GES-1 cells, the ABI1 expression in AGS cells was lowered significantly. There were no significant differences in the ABI1 mRNA and protein expression between the AGS and AGS- MSCV-GFP groups. Compared to those of the AGS group, the ABI1 mRNA expression levels of the AGS- MSCV-GFP-ABI1 group increased by 1.87 times （ P = 0. 002 ）. The protein expression levels of the AGS- MSCV-GFP-ABI1 group were remarkably higher than those of the AGS and AGS-MSCV-GFP groups （P = 0. 002）. CCK-8 assay showed that there were no significant differences in the proliferation rates at different time points between the AGS and AGS-MSCV-GFP groups. However, the proliferation rates at the time points of 24, 48, 72 and 96 hours of the AGS-MSCV-GFP-ABI1 were 1.46 ± 0. 31, 4. 75 ± O. 12, 6. 62 ± 0. 32 and 8.96 ± 0. 27 respectively. And they were significantly lower than the proliferation rates of the AGS and AGSMSCV-GFP groups （ P 〈 0. 01 ）. Conclusion ABI1 gene is down-regulated in gastric cancer cells. The ABI1 overexpression effectively inhibits the proliferation in human gastric cancer cell lines. It suggests that ABI1 may be involved in gastric cancer pathogenesis by regulating the proliferat
出处
《中华医学杂志》
CAS
CSCD
北大核心
2009年第44期3111-3115,共5页
National Medical Journal of China
关键词
胃肿瘤
细胞增殖
Abl相互作用蛋白1
Stomach neoplasms
Cell proliferation
Abl-interacting protein 1
