中文摘要：

目的在体外共培养体系中研究胆管癌细胞对内皮细胞的作用。方法建立胆管癌QBC939细胞与内皮细胞的体外共培养体系设为共培养组，同期单独培养的内皮细胞设为内皮细胞组，同样数量的内皮细胞和胆管癌细胞分别单独培养的上清液混合液设为混合组。对内皮细胞进行光镜及扫描电镜观察，并用免疫荧光法检测内皮细胞蛋白水解酶pp125焦点激酶（pp125FAK）、基质金属蛋白酶2和9（MMP-2、MMP-9）、人尿激酶型纤维蛋白溶解酶原激活物（uPA）表达的变化，明胶酶谱法测定内皮细胞MMP-2及MMP-9的活性。采用配对t检验分析其结果。结果与内皮细胞组比较，共培养组内皮细胞之间的间隙增大。内皮细胞组pp125FAK、MMP-2、MMP-9、uPA的平均荧光强度分别为394±51、455±82、377±48、422±55，而共培养组表达增强，分别为1096±128、931±72、815±76、801±56，两组比较差异有统计学意义（t=6．53，4．32，3．61，3．45，P〈0．05）。混合组MMP-2、MMP-9灰度扫描值分别为240．2±15．2和2．4±0．8，共培养组的MMP-2、MMP-9分别为687．4±43．6和150．9±13．2，两组比较差异有统计学意义（t=4．89，5．43，P〈0．05）。结论共培养后的内皮细胞之间缝隙增大，蛋白水解酶表达增强，它可能参与降解内皮细胞外基质，促进了胆管癌的转移。

英文摘要：

Objective To study the influence of cholangiocarcinoma cells on endothelial cells in a co-culture system. Methods A co-cuhure system of cholangiocarcinoma cell line QBC939 and endothelial cells was established in vitro （co-culture group）. Endothelial cells were cultured individually during the same time （control group）. The mixed supematant of cholangiocarcinoma cells and endothelial cells was in the mixed group. Light microscopy and transmission electron microscopy were used to observe the morphology of the endothelial cells. Changes in expression of pp125FAK, MMP-2, MMP-9 and uPA of the endothelial cells were detected by immunofluorescence, and the activities of MMP-2 and MMP-9 were detected by gelatin zymography. All the data were analyzed by paired t test. Results The intercellular space between endothelial cells in co-culture group was wider than in the control group. The expression of pp125FAK, MMP-2, MMP-9 and uPA was 394 ± 51,455 ±82, 377 ±48, 422 ±55 in control group, and was 1096 ± 128, 931 ±72, 815 ±76, 801±56 in the co-culture group. The difference between the 2 groups had statistical significance （ t = 6.53, 4.32, 3.61, 3.45, P 〈 0.05）. The values of gray-scale scanning of MMP-2 and MMP-9 in the mixed group were 240.2 ± 15.2 and 2.4 ±0.8, respectively. The values of gray-scale scanning of MMP-2 and MMP-9 in the co-cuhure group were significantly increased, they were 687.4 ± 43.6 and 150.9 ± 13.2, respectively （ t = 4.89, 5.43, P 〈 0.05 ）. Conclusions The intercellular space between endothelial cells and the expression of the proteolytic enzymes are increased after co-culturing endothelial cells with cholangiocareinoma cells. Proteolytic enzymes may be involved in the process of degradation of subendothelial matrix, and promotes the metastasis of cholangiocarcinoma.