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.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2012年第4期232-235,共4页
Chinese Journal of Digestion
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
Recombinant proteins
Stomach neoplasms
Tumor cells, cultured
Vascular endothelial growth factors
Hypoxia-inducible factor 1, alpha subunit
Anoxia
作者简介
通信作者：任建林，Email：renjianl@xmu．edu．cn
