摘要
目的探讨阻断胃泌素受体对胃癌细胞增殖、凋亡及其相关通路中关键蛋白表达的影响。方法试验组使用终浓度为5mmol/L的丙谷胺(胃泌素受体阻断剂)处理胃癌细胞SGC-7901和AGS 6d,以不加丙谷胺培养的胃癌细胞SGC-7901和AGS为对照组。四甲基偶氮唑蓝比色(MTT)法检测各组细胞的生长并绘制细胞生长曲线;流式细胞术检测各组细胞的细胞周期,Annexin V-FITC/PI双染法检测各组细胞的凋亡;实时荧光定量PCR(RT-qPCR)方法检测典型Wnt/β-catenin通路、核因子κB、磷脂酰肌醇3激酶-丝氨酸-哺乳动物雷帕霉素靶蛋白中关键蛋白β-连环蛋白(β-catenin)、核转录因子RelA、哺乳动物雷帕霉素靶蛋白、糖原合酶激酶3βmRNAs的表达;免疫蛋白印迹检测β-catenin蛋白质的表达。结果用丙谷胺处理后,试验组细胞的生长速度低于对照组细胞,细胞周期中S期细胞百分数也低于对照组细胞,而G_0/G_1期细胞百分数高于对照组细胞(P<0.05);试验组的细胞凋亡数高于对照组(P<0.05);RT-qPCR结果显示:丙谷胺处理后,胃癌细胞中β-catenin的mRNA表达量降低(P<0.05)。Western blot结果显示丙谷胺处理后,β-catenin蛋白质的表达量降低(P<0.05)。结论阻断胃泌素受体能下调胃癌细胞中β-catenin的表达,抑制细胞增殖,同时促进细胞凋亡。
Objective To investigate the effects of blocking gastrin receptor on the proliferation,apoptosis and expression of key proteins in the related pathway in gastric cancer cell lines.Methods In the experimental group,the gastric cancer cell lines SGC-7901 and AGS cells were treated with 5mmol/L proglumide,a kind of a gastrin receptor antagonist.And the normal cultured gastric cancer cells SGC-7901 and AGS were used in control group.The growth of each group was detected by MTT assay;the cell growth curve was drawn by flow cytometry;the cell cycle of each group was detected by flow cytometry.Annexin V-FITC/PI double staining was used to detect the cell growth of apoptosis.The relative mRNA expression ofβ-catenin,nuclear factor-P65,mammalian target of rapamycin and glycogen synthase kinase 3beta in Wnt,NF-κB and PI3K-AKT-MTOR pathways were detected by RTqPCR.The expression ofβ-catenin protein was detected by Western blotting.Results After treatment with proglumide,the growth of the cells in the experimental group was lower than that in the control group;and the proportion of S phase cells in the cell cycle was also lower than that in the control group,but the proportion of cells in G_0/G_1 phase was higher than that in the control group（P〈0.05）.The percentage of apoptotic cells was also increased after treatment with proglumide（P〈0.05）.Furthermore,proglumide treatment significantly reduced the expression ofβ-catenin at both mRNA and protein levels（P〈0.05）.Conclusion Blocking gastrin receptor can down-regulate the expression ofβ-catenin,inhibit the cell proliferation and promote the cell apoptosis in gastric cancer cells.
出处
《重庆医学》
CAS
北大核心
2017年第15期2017-2020,共4页
Chongqing medicine
基金
国家自然科学基金资助项目(81260303
31560326)
贵州省科技合作计划项目(黔科合LH字[2015]7360)
