体外共培养体系中胆管癌细胞对人脐静脉内皮细胞b-FGF和VEGF表达的影响  

Influence of cholangiocarcinoma cell lines on expression of b-FGF and VEGF in HUVEC in a co-culture system

作　　者：李薇[1] 杨芸[1] 宋富强[1] 冯亚星[1] 李昆[1] 

机构地区：[1]成都军区总医院临床实验医学研究与保障中心,成都610083

出　　处：《重庆医学》2015年第13期1749-1751,共3页Chongqing medicine

基　　金：四川省卫生厅科学研究项目(100091)

摘　　要：目的探讨体外共培养体系中胆管癌细胞(QBC939)对人脐静脉内皮细胞(HUVEC)、碱性成纤维细胞生长因子(bFGF)和血管内皮生长因子(VEGF)表达的影响。方法建立胆管癌QBC939细胞与HUVEC体外共培养体系,采用CCK-8法检测共培养不同时相HUVEC的增殖;ELISA检测共培养上清液中b-FGF含量;qRT-PCR和Western blot检测共培养不同时相点HUVEC细胞VEGF mRNA和蛋白质的表达。结果共培养后12、24、48h和72h,HUVEC增殖较对照组显著增高(P<0.01)。与0h组比较,共培养12、24、48h上清液中b-FGF含量显著增高(P<0.05)。共培养12h时VEGF mRNA表达显著增加,并持续至72h(P<0.01),VEGF蛋白表达在共培养12h后显著增高,24、48、72h时与对照组比较差异有统计学意义(P<0.05)。结论人胆管癌QBC939细胞可促进血管内皮细胞增殖、增加b-FGF分泌和VEGF表达。b-FGF和VEGF表达增加与肿瘤血管新生,以及肿瘤的生长和转移可能具有相关性。Objective To study the influence of cholangiocarcinoma cell lines on the expression of b-FGF and VEGF in hu- man umbilical vein endothelial cell （HUVEC） in a co-culture system. Methods A co-culture system of eholangioearcinoma cell lines QBC939 and HUVEC were established in vitro. QBC939 cells and HUVCE were co-cultured for 12,24,48 and 72 h respectively,the proliferation of HUVEC was detected by CCK-8 assay. The b-FGF level in cell culture supernatant was detected by ELISA assay. The mRNA and protein expression levels of VEGF were detected by qRT-PCR and Western blot analysis. Results The prolifera- tion of HUVEC was significantly increased at 12,24,48 and 72 h after incubation in a co-culture system （P〈0.01）. The mRNA expression level of VEGF in t-IUVEC was gradually increased after 12 h incubation with a time-dependent manner. It showed that the protein level of b-FGF and VEGF was also significantly increased at 12,24,48 h after incubation as compared to the control（P 0.05）. Conclusion Human cholangiocarcinoma cells can promote the proliferation of endothelial cells,increase the secretion of b- FGF and upregulate the expression of VEGF, suggesting that the expressions of b-FGF and VEGF may be correlated with the tumor angiogenesis.

关 键 词：胆管肿瘤 内皮细胞 脐静脉 血管内皮生长因子A 

分 类 号：R735.8[医药卫生—肿瘤]