缺氧诱导胃癌细胞SGC-7901中三叶因子3与血管内皮生长因子相互关系研究  

The study on the relationship between trefoil factor family 3 and vascular endothelial growth factor in hypoxic induced gastric cancer SGC-7901 cells

作　　者：黄庆文[1] 韩佳[1] 王琳[1] 叶震世[1] 巴亚斯古楞[1] 任建林[1] 

机构地区：[1]厦门大学附属中山医院消化科厦门大学消化疾病研究所厦门市消化疾病中心,福建厦门361004

出　　处：《中华消化杂志》2012年第4期232-235,共4页Chinese Journal of Digestion

摘　　要：目的探讨在缺氧条件下胃癌细胞SGC-7901中三叶因子3（TFF3）与血管内皮生长因子（VEGF）及缺氧诱导因子（HIF—1a）的相互关系，了解TFF3在胃癌发生发展过程中的作用机制。方法使用氯化钴（CoCl2）构建胃癌细胞株SGC-7901的缺氧模型。运用携带靶向干扰人类TFF3的pU6-siTFF3和pU6-mock分别转染胃癌细胞株SGC-7901，以嘌呤霉素为筛选药物，建立稳定特异性抑制TFF3的胃癌细胞株。在缺氧环境和常氧环境下培养胃癌细胞株SGC-7901和靶向干扰TFF3后的胃癌细胞株SGC-7901，运用定量PCR、ELISA和Western印迹分析等方法分别测定其TFF3、HIF-1a和VEGF的蛋白和mRNA表达情况。运用免疫荧光法观察在缺氧环境和常氧环境下胃癌细胞株SGC-7901中TFF3、HIF-1a的分布及表达量。结果CoCl2缺氧处理能诱导胃癌细胞株SGC-7901中HIF-1d、TFF3和VEGFmRNA表达量上升（分别为33．4±1．8、14．8±1．1和15．1±1．2）。稳定干扰TFF3的SGC-7901细胞在缺氧诱导下能下调VEGF和HIF-1蛋白的表达。结论TFF3介导调节了缺氧条件下VEGF和HIF-1的表达，TFF3有可能是潜在的抗血管生成胃癌治疗靶点。Objective To explore the relationship of trefoil factor family 3 （TFF3）, vascular endothelial growth factor （VEGF） and hypoxianducible factor （HIF）-1a in gastric cancer SGC-7901 cells under hypoxic condition and try to investigate the mechanism of TFF3 in the genesis and development of gastric cancer. Methods The hypoxic model of gastric cancer SGC-7901 cell was induced by COCl2. Gastric cancer cell line SGC-7901 cells were transfected with pU6-siTFF3 plasmid which carrying RNAi targeted to human TFF3 and pU6-mock. Puromycin was selected as screening medicine. The stable and specific TFF3 inhibited gastric cancer ceil line was established. Gastric cancer cell line SGC-7901 and TFF3 RNAi targeted gastric cancer cell line SGC-7901 were cultured under hypoxic condition and normoxic condition. The expression of TFF3, VEGF and HIF-1a at protein and mRNA level were detected by RT-PCR, Western blot and ELISA assay. The distribution and expression of TFF3 and HIF-1a in gastric cancer cell line SGC-7901 cells under normoxia and hypoxic condition were determined with immunofluorescence staining. Results The expressions of HIF-1a, TFF3 and VEGF in gastric cancer SGC-7901 cell increased under COCl2 induced hypoxic condition （33.4 ± 1. 8, 14. 8 ± 1. 1 and 15. 1 ± 1. 2, respectively）. Under hypoxic condition, the expression of VEGF and HIF-1a protein reduced in stable TFF3 RNAi SGC-7901 cells. Conclusion TFF3 mediated the regulation of VEGF and HIF-1a expression under hypoxic condition. TFF3 might be a potential anti-angiogenic target in gastric cancer treatment.

关 键 词：重组蛋白质类 胃肿瘤 肿瘤细胞 培养的 血管表皮生长因子类 缺氧诱导因子 1 a亚基 缺氧 

分 类 号：R735.2[医药卫生—肿瘤]