摘要
目的 探讨17肽胃泌素(Gas-17)及其受体拮抗剂对人胃癌细胞株MKN45增殖及三叶因子1(TTF1)、三叶因子3(TFF3)表达的影响,并分析TTF1、TFF3在胃癌病变过程中的作用。方法培养人胃癌细胞株MKN45后按药物干预分组:第1组:17肽胃泌素组,培养液中Gas-17终浓度为1~1000 nmo1/L;第2组:丙谷胺(PGL)组,培养液中PGL终浓度为0.1~10 mmol/L;第3组:17肽胃泌素+丙谷胺组(联合用药组),培养液中Gas-17终浓度为100 nmo1/L、PGL终浓度为1~10 mmol/L;以不加药培养液为对照组。采用四甲基偶氮唑盐(MTT)比色法观察各组细胞增殖活力。蛋白免疫印迹法(Western blot)测定17肽胃泌素组(培养液中Gas-17终浓度为1~100 nmo1/L),丙谷胺组(PGL终浓度为10mmol/L),联合用药组(Gas-17终浓度为100 nmo1/L、PGL终浓度为10 mmol/L)及不加药对照组中TFF1和TFF3蛋白的表达变化。结果 MTT结果显示:Gas-17在1~1000 nmo1/L时具有明显的促MKN45细胞增殖作用(P【0.05);PGL在1~10 mmol/L时有显著的抑制MKN45细胞增殖作用(P【0.05);Gas-17+PGL组中,PGL(1~10 mmol/L)能阻断并抑制Gas-17对胃癌细胞MKN45的促增殖作用(P【0.05)。Western blot结果显示:在Gas-17组中TFF1蛋白表达减弱(P【0.05),而TFF3蛋白表达增强(P【0.05);PGL组中TFF1蛋白表达增强而TFF3蛋白表达减弱;Gas-17+PGL组中,PGL能阻断Gas-17诱导的TFF1蛋白表达下调(P【0.05),阻断Gas-17诱导的TFF3蛋白表达上调(P【0.05)。结论Gas-17可诱导人胃癌细胞MKN45增殖,其受体抑制剂PGL能阻断并抑制这一作用。胃癌细胞株MKN45中有TFF1和TTF3蛋白的表达,Gas-17促进TFF1蛋白表达下调,而促进TFF3蛋白表达上调,这可能是胃泌素诱导胃癌发生发展的的机制之一。
Objective To explore the effects of gastrin-17 and its antagonist PGL on cell proliferation and the expression of trefoil factor1(TFF1), trefoil factor3(TFF3) in human gastric cancer line MKN-45, and to analyze the function of TFF1 and TFF3 in the development of gastric lesions. Methods MKN45 cells were incubated in the medium with Gas-17(1~1000 nmo1/L), proglumide(1~10 mmol/L) and the combination of these two agents(100nmo1/L Gas-17 and 1~10 mmol/L proglumide). Medium with no drug was used as control. Cell growth and proliferation of MKN45 were analyzed with MTT assay. The expression of TFF1 and TFF3 were determined by Western blot in MKN45 cells which were incubated with Gas-17(1~100nmo1/L), proglumide(10 mmol/L), the combination(100nmo1/L Gas-17 and 10mmol/L proglumide) and the control group. Results MTT showed Gas-17 significantly promoted cell proliferation at the concentration of 1~1000 nmo1/L(P<0.05); PGL significantly inhibited cell proliferation at the concentration of 1~10mmol/L(P<0.05); In the combination group, PGL(1~10mmol/L) significantly blocked and inhibited cell proliferation induced by Gas-17(P<0.05). Meanwhile, Western blot showed the expression of TFF1 and TFF3 protein was found in gastric cancer cell line MKN45; Gas-17 significantly increased TFF3 while inhibited TFF1(P<0.05); PGL significantly up-regulated TFF1 while inhibited TFF3. In the combination group, PGL significantly inhibited the down-regulation of TFF1 and up-regulation of TFF3 stimulated by Gas-17(P<0.05). Conclusion Gas-17 would induce cell proliferation in human gastric cancer line MKN45 while its antagonist PGL could significantly block and inhibit this effect. Gas-17 could inhibit TFF1 and increase TFF3 which may be one of the mechanisms of Gas-17 inducing the development of gastric carcinoma.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2014年第6期545-548,共4页
Cancer Research on Prevention and Treatment
基金
贵州省科技厅社会攻关计划资助项目(SY[2010]3085号)
