P38信号转导通路在胃泌素促进人大肠癌细胞SW480增殖中的作用及其机制  

Role of P38 signaling pathway and its regulation mechanisms in gastrin-induced cell proliferation and apoptosis of human large intestinal cancer cell line SW480

作　　者：茆家定[1] 陶亮[1] 吴佩[1] 

机构地区：[1]皖南医学院附属弋矶山医院胃肠二科,安徽芜湖241001

出　　处：《皖南医学院学报》2014年第5期389-392,共4页Journal of Wannan Medical College

基　　金：安徽省自然科学研究基金项目(11040606M182);安徽省教育厅自然科学研究基金项目(KJ2010B242)

摘　　要：目的：探讨胃泌素对人大肠癌细胞株SW480增殖、凋亡的影响，以及P38-丝裂素活化蛋白激酶（ MAPK）信号转导通路的关系。方法：体外培养大肠癌SW480细胞株，将细胞分为对照组、胃泌素组、丙谷胺组和胃泌素＋丙谷胺组，对照组不加药，其余组加不同浓度的药物处理。运用MTT法观察细胞凋亡情况，确立5-肽胃泌素和丙谷胺处理SW480细胞的最佳浓度；流式细胞术检测各组细胞增殖指数的变化；qRT-PCR检测大肠癌SW480细胞株胃泌素受体／胆囊收缩素-B受体（ CCK-2R） mRNA表达情况；Western blot 检测P38蛋白表达及其磷酸化水平。采用方差分析和SNK-q检验进行统计学分析。结果：SW480细胞中存在CCK-2R mRNA表达。胃泌素在6.25～200.00 mg／L范围内能抑制大肠癌SW480细胞的凋亡，且当浓度达12.50 mg／L时为最佳浓度（ P＜0.05）；单独丙谷胺对大肠癌SW480细胞凋亡无明显影响（ P＞0.05），但丙谷胺在8.00～256.00 mg／L范围内能明显拮抗胃泌素抑制SW480细胞的凋亡作用，其最佳浓度为16.00 mg／L （ P＜0.05）。胃泌素（12.50 mg／L）组细胞增殖指数为（34.56±1.41）％，显著高于对照组的（28.74±1.61）％和胃泌素（12.50 mg／L）＋丙谷胺（16.00 mg／L）组的（28.72±1.78）％（P＜0.05），而丙谷胺（16.00 mg／L）组细胞增殖指数为（27.75±2.29）％，与对照组比较差异无统计学意义（P＞0.05）；P38蛋白及其磷酸化在胃泌素组（12.50 mg／L）中表达水平低于对照组（P＜0.01），也低于胃泌素（12.50 mg／L）＋丙谷胺（16.00 mg／L）组（ P＜0.01）。结论：胃泌素可以抑制大肠癌细胞株SW480的凋亡、促进增殖，其作用可能是通过抑制P38及其磷酸化水平来实现的，但能够被其受体拮抗剂丙谷胺抑制。Objective：To investigate the effects of gastrin on the proliferation and apoptosis of human large intestinal carcinoma cell line SW 480 and the relationship of signal transduction pathway in p38 mitogen-activated protein kinases（P38-MAPK）.Methods：SW 480 cell line of human large intestinal cancer was in vitro incubated and divided into groups of control ,gastrin use,proglumide intervention and gastrin ＋proglumide.The control group were free of medication,and the remaining groups were treated with corresponding drugs in different dosage .MTT reduction assay was performed to detect the cell ap-optosis status and determine the optimal dosage of pentagstrin and proglumide affecting SW480 cell line proliferation.Flow cytometry was used to estimate the proliferation index in each group,and qRT-PCR was performed to detect the expression of gastrin receptor （CCK-2R） mRNA in SW480 cells.Western blot was performed to examine the expression of P38 protein and its phosphorylation levels.All results were analyzed with χ2 test and SNK-q test.Results：qRT-PCR detection showed that CCK-2R mRNA was expressed in the SW480 cell line.Pentagastrin inhibited SW480 cell apoptosis in a dose ranging from 6.25 to 200 mg/L,and the optimal dosage was 12.50 mg/L（P〈0.05）.Proglumide was capable of inhibiting the cell proliferation in dosage of 8.00 -256.00 mg/L,and the optimal effect was in dose of 16.00 mg/L（P〈0.05）.The proliferation index in pentagastrin group was （34.56 ±1.41）%,which was significantly higher than that of control group（28.74 ±1.61） % and combined treatment group（28.72 ±1.78）%（P〈0.05）,and the index differ-ence was not significant between proglumide group and the controls（P〉0.05）.P38 protein expression and its phosphorylation level were lower in penta-gastrin group as compared with the controls and combination treatment group（P〈0.01）.Conclusion：Gastrin can inhibit the apoptosis of human large in-testinal cancer cell line SW480 and improve the cell proliferation.This

关 键 词：结肠癌 胃泌素 丙谷胺 P38 

分 类 号：R735.34[医药卫生—肿瘤]