CAJ | 학술논문

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-1α)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-1α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-1α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H1F-1α、TFF3和VEGF mRNA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.
목적 탐토재결양조건하위암세포SGC-7901중삼협인자3(TFF3)여혈관내피생장인자(VEGF)급결양유도인자(HIF-1α)적상호관계,료해TFF3재위암발생발전과정중적작용궤제.방법 사용록화고( CoCl2)구건위암세포주SGC-7901적결양모형.운용휴대파향간우인류TFF3적pU6 siTFF3화pU6-mock분별전염위암세포주SGC-7901.이표령매소위사선약물,건립은정특이성억제TFF3적위암세포주.재결양배경화상양배경하배양위암세포주SGC-7901화파향간우TFF3후적위암세포주SGC-7901,운용정량PCR、ELISA화Western인적분석등방법분별측정기TFF3、HIF-1α화VEGF적단백화mRNA표체정황.운용면역형광법관찰재결양배경화상양배경하위암세포주SGC-7901중TFF3、HIF-1α적분포급표체량.결과 CoCl2결양처리능유도위암세포주SGC-7901중H1F-1α、TFF3화VEGF mRNA표체량상승(분별위33.4±1 8、14.8±1.1화15.1±1.2).은정간우TFF3적SGC-7901세포재결양유도하능하조VEGF화HIF-1단백적표체.결론 TFF3개도조절료결양조건하VEGF화HIF-1적표체,TFF3유가능시잠재적항혈관생성위암치료파점.
Objective To explore the relationship of trefoil factor family 3 (TFF3),vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF)-1α 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 transfccted 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 cell 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-1α in gastric cancer cell line SGC-7901 cells uuder 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 hypoxie condition,the expression of VEGF and HIF-1α protein reduced in stable TFF3 RNAi SGC-7901cells.Conclusion TFF3 mediated the regulation of VEGF and HIF-1α expression under hypoxic condition.TFF3 might be a potential anti-angiogenic target in gastric cancer treatment.