胆囊收缩素对人胆管癌QBC_(939)细胞分泌MMPs、TIMPs水平的影响  

Effect of cholecystokinin on expression of MMPs and TIMPs in human cholangiocarcinoma QBC_(939) cells

作　　者：杨星[1] 张丰深[1] 孙海[1] 邓小明[1] 陈焱[1] 

机构地区：[1]解放军第324医院肝胆外科,重庆400020

出　　处：《第三军医大学学报》2014年第16期1684-1688,共5页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2010BB5159)~~

摘　　要：目的观察胆囊收缩素（cholecystokinin,CCK）对人胆管癌QBC939细胞基质金属蛋白酶（mtrix metalloproteinases,MMPs）及其内源性组织抑制剂（tissue inhibitors of matrix metalloproteinases,TIMPs）表达的影响,阐明CCK促进胆管癌周围神经浸润（perineural invasion,PNI）可能的分子机制。方法将人胆管癌QBC939细胞分为对照组与实验组。对照组为未经处理的胆管癌细胞QBC939;实验1~8组分别加入10-13、10-11、10-9、10-7mol/L CCK8s,10-7mol/L L18（CCK-A受体拮抗剂）、10-7mol/L L60（CCK-B受体拮抗剂）、10-7mol/L CCK8s＋10-7mol/L L18、10-7mol/L CCK8s＋10-7mol/L L60的QBC939,孵育48 h,采用Transwell细胞侵袭实验比较各组QBC939细胞侵袭能力的差异,并用Real-time PCR和Western blot检测各组MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA和蛋白表达情况。各组间比较采用方差分析,采用Spearman法进行相关性检验。结果 CCK8s能够明显促进QBC939细胞的侵袭能力,L18及L60均能明显减弱CCK8s对QBC939细胞的作用。与对照组相比,实验1~4组MMP-2及MMP-9 mRNA和蛋白的表达水平明显升高（P〈0.01）,TIMP-1及TIMP-2mRNA和蛋白的表达水平明显降低（P〈0.01）;实验5~6组MMP-2及MMP-9 mRNA和蛋白的表达水平明显降低（P〈0.01）,TIMP-1及TIMP-2 mRNA和蛋白的表达水平明显升高（P〈0.01）;实验7~8组MMPs及TIMPs mRNA和蛋白表达水平与对照组比较差异无统计学意义（P〉0.05）。实验1~4组MMP-2 mRNA和蛋白的表达水平与CCK浓度呈正相关（相关系数r分别为0.972、0.963,P〈0.01）,而TIMP-2 mRNA和蛋白的表达水平与CCK浓度呈负相关（相关系数r分别为-0.974、-0.952,P〈0.01）。MMP-9、TIMP-1的变化与CCK浓度间无明显相关性;实验组5与实验组6比较、实验组7与实验组8比较,MMP-2、MMP-9、TIMP-1及TIMP-2 mRNA和蛋白表达水平差异无统计学意义（P〉0.05）。结论 CCK能明显促进QBC939细胞侵袭能力及MMPs的表达、降低TIMPs的表达,可能主要通�Objective To investigate the effect of cholecystokinin （CCK） on the expression of matrix metalloproteinases （MMPs） and tissue inhibitors of matrix metalloproteinases （TIMPs） in human cholangiocarcinoma QBC939 cells. Methods Human cholangiocarcinoma QBC939 cells were assigned into a control group and 8 experimental groups. The experimental groups （group 1 to group 8） were treated with 10-13, 10-11, 10-9, 10-7 and 10-7mol/L L18 （a CCK-A receptor antagonist）, 10-7 mol/L L60 （a CCK-B receptor antagonist）, 10-7 mol/L CCK8s ＋ 10-7 mol/L L18, and 10-7 mol/L CCK8s ＋ 10-7 mol/L L60, respectively, for 48 h. Transwell Matrigel invasion assay was used to study the impact on the invasion ability of QBC939 cells. The mRNA and protein expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 were detected by real-time PCR and Western blotting. All the data were analyzed using the analysis of variance and Spearman rank correlation. Results CCK8s could promote invasion ability of QBC939 cells significantly. The mRNA and protein expression of MMP-2 and MMP-9 in experimental groups 1 to 4 were significantly higher than those of the control group （P〈0.01）, and the mRNA and protein expression of TIMP-1 and TIMP-2 were significantly lower （P〈0.01）. The mRNA and protein expressions of MMP-2 and MMP-9 in experimental groups 5 and 6 were obviously lower than those of the control group （P〈0.01）, and the expression of TIMP-1 and TIMP-2 were obviously higher （P〈0.01）. There was no significant difference in the mRNA and protein expression of MMPs and TIMPs among experimental groups 7 and 8 and the control group. The mRNA and protein expression of MMP-2 in experimental groups 1 to 4 were positively correlated with the concentrations of CCK （r=0.972, 0.963; P〈0.01）, while those of TIMP-2 were negatively correlated with the concentrations of CCK （r=-0.974, -0.952; P〈0.01）. Conclusion CCK can significantly promote the invasion ability of QBC939 cells an

关 键 词：胆囊收缩素 基质金属蛋白酶 胆管癌 周围神经浸润 

分 类 号：R345[医药卫生—基础医学] R735.8