Cui Mei-hua, Yu Wei-dong, Dong Jian-qiang, Liu Yu-lan
Department of Gastroenterlogy, Peking University People's Hospital, Beijing 100044, China.
Zhonghua Yi Xue Za Zhi. 2009 Dec 1;89(44):3111-5.
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.
Firstly the ABI1 expression in GES-1 and AGS cells were identified by immunohistochemistry, immunofluorescence, 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.
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 +/- 0.12, 6.62 +/- 0.32 and 8.96 +/- 0.27 respectively. And they were significantly lower than the proliferation rates of the AGS and AGS-MSCV-GFP groups (P < 0.01).
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 proliferation of gastric carcinomas cells.
研究Abl相互作用蛋白1（ABI1）在正常胃黏膜细胞系GES-1和胃癌细胞系AGS中的表达情况，以及ABI1基因过表达对人胃癌细胞AGS体外增殖的影响。
首先采用免疫组织化学、免疫荧光、RT-PCR、实时荧光定量PCR及蛋白质免疫印迹法检测GES-1和AGS细胞中ABI1的表达。其次培养人胃癌细胞系AGS，并用重组MSCV-GFP-ABI1质粒或空白质粒MSCV-GPF进行转染。采用实时荧光定量PCR和蛋白质免疫印迹法检测ABI1的mRNA和蛋白表达。最后通过CCK-8法检测细胞增殖情况。
ABI1在正常胃黏膜细胞系GES-1和胃癌细胞系AGS中均有表达。与GES-1细胞相比，AGS细胞中ABI1的表达明显降低。AGS组与AGS-MSCV-GFP组之间ABI1的mRNA和蛋白表达无显著差异。与AGS组相比，AGS-MSCV-GFP-ABI1组的ABI1 mRNA表达水平增加了1.87倍（P = 0.002）。AGS-MSCV-GFP-ABI1组的蛋白表达水平显著高于AGS组和AGS-MSCV-GFP组（P = 0.002）。CCK-8法检测结果显示，AGS组与AGS-MSCV-GFP组在不同时间点的增殖率无显著差异。然而，AGS-MSCV-GFP-ABI1组在24、48、72和96小时的增殖率分别为1.46±0.31、4.75±0.12、6.62±0.32和8.96±0.27。且均显著低于AGS组和AGS-MSCV-GFP组（P < 0.01）。
ABI1基因在胃癌细胞中表达下调。ABI1过表达可有效抑制人胃癌细胞系的增殖。提示ABI1可能通过调控胃癌细胞的增殖参与胃癌的发病机制。
