摘要
目的:探讨HIF-1α沉默后对食管癌细胞株TE13的生物学行为的影响.方法:应用倒置荧光显微镜观察HIF-1α的干扰质粒转染食管癌细胞TE13后绿色荧光蛋白的表达;采用Western blot方法检测HIF-1α蛋白的表达;四甲基偶氮唑蓝(MTT)法检测转染前后细胞增殖能力的变化;Transwell方法检测干扰前后细胞迁移能力的变化;流式细胞术检测干扰前后细胞周期的变化.结果:TE13/12单克隆干扰效果好,Western blot结果显示无HIF-1α表达.HIF-1α被干扰后,细胞增殖能力明显减弱(P<0.05),运动迁移能力显著下降,与未转染的细胞相比穿过人工基底膜的细胞数明显减少(18.2±3.7vs103.8±8.5,P<0.05).与未转染组相比,TE13/12的细胞周期发生变化,G2/M期细胞明显减少(5.99%±1.19%vs20.47%±4.30%,P<0.05),S期增加(64.67%±1.98%vs48.53%±3.89%,P<0.05).结论:RNA干扰可引起TE13中HIF-1α的沉默,而HIF-1α表达下调后食管癌细胞株TE13的增殖与迁移能力均减弱.推测阻断HIF-1α通路有可能成为治疗人食管鳞癌的新靶点.
AIM：To investigate the impact of RNA interference-mediated downregulation of hypoxia-inducible factor-1α （HIF-1α） on the biological behavior of esophageal squamous cell carcinoma TE13 cells. METHODS：The plasmid harboring small interfering RNA targeting HIF-1α was introduced into TE13 cells by liposome-mediated transfection. The expression of HIF-1α protein was measured by Western blot. The proliferation of TE13 cells was determined by methyl thiazolyl tetrazolium （MTT） colorimetric assay. The migration of TE13 cells was evaluated by Transwell migration assay. Cell cycle analysis was performed by fluorescence-activated cell sorting （FACS）. RESULTS：HIF-1α expression was markedly downregulated in TE13/12 cell clone （transfected with the plasmid harboring small interfering RNA targeting HIF-1α）. The expression of HIF-1α protein in TE13/12 cells could not be detected by Western blot. The proliferation of TE13/12 cells decreased signifi cantly compared with untransfected TE13 cells （P〈0.05）. Transwell migration assay showed that less TE13/12 cells could move through the artif icial basement membrane when compared with untransfected TE13 cells （18.2 ± 3.7 vs 103.8 ± 8.5,P〈0.05）. The cell cycle of TE13/12 cells was changed. Compared with untransfected TE13 cells,the percentage of TE13/12 cells in G2/M phase decreased significantly （5.99% ± 1.19% vs 20.47% ± 4.30%,P〈0.05）,whereas the percentage of TE13/12 cells in S phase increased remarkably （64.67% ± 1.98% vs 48.53% ± 3.89%,P〈0.05）. CONCLUSION：RNA interference-mediated downregulation of HIF-1α can attenuate the proliferation and migration of TE13 cells. Blockade of the HIF-1α pathway may provide a new approach to the treatment of human esophagus squamous cell carcinoma.
出处
《世界华人消化杂志》
CAS
北大核心
2009年第27期2762-2766,共5页
World Chinese Journal of Digestology
关键词
食管鳞癌
缺氧诱导因子-1Α
RNA干扰
Esophageal squamous cell carcinoma Hypoxia-inducible factor-1α RNA interference
