文章摘要
曾楷峰,施瑞华,朱〓宏,张国新,郝〓波.RNA干扰沉默缺氧诱导因子-1α对胃癌细胞生物学行为的影响[J].南京医科大学学报,2010,(2):144~149
RNA干扰沉默缺氧诱导因子-1α对胃癌细胞生物学行为的影响
Effects of RNA interference targeting HIF-1α gene on the biological behavior of gastric cancer cells
投稿时间:2009-10-25  
DOI:10.7655
中文关键词: 胃癌  缺氧诱导因子-1a  RNA干扰  侵袭
英文关键词: gastric cancer  RNA interference  hypoxia inducible factor-1α  invasion
基金项目:国家自然科学基金资助项目(30770991)
作者单位
曾楷峰 南京医科大学第一附属医院消化科,江苏 南京〓210029 
施瑞华  
朱〓宏  
张国新  
郝〓波  
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中文摘要:
      目的:观察RNA干扰沉默缺氧诱导因子-1α(HIF-1α)对胃癌细胞增殖﹑凋亡﹑侵袭性的影响并探讨其可能的分子机制?方法:将HIF-1α短发夹RNA(shRNA)重组质粒pGCsi-shHIF-1α稳定转染人胃癌细胞SGC-7901;采用RT-PCR和Western blot法检测HIF-1α mRNA?蛋白表达水平,Western blot法检测葡萄糖转运蛋白1(Glut-1)?基质金属蛋白酶2(MMP-2)及趋化因子受体CXCR4蛋白表达;MTT法?流式细胞仪?transwell试验分别检测转染细胞的增殖?凋亡及侵袭迁移能力?结果:稳定转染pGCsi-shHIF-1α后,SGC-7901细胞HIF-1α表达在mRNA及蛋白水平均受到明显抑制,Glut-1﹑MMP-2﹑CXCR4蛋白表达较对照组降低;细胞增殖能力较对照组明显降低,细胞凋亡率明显高于对照组(P < 0.05);细胞体外侵袭迁移能力受到明显抑制(P < 0.01)?结论:pGCsi-shHIF-1α能有效沉默胃癌细胞SGC-7901 HIF-1α基因表达,抑制胃癌细胞增殖?迁移及侵袭并诱导其凋亡,提示HIF-1α可能通过上调Glut-1﹑MMP-2和CXCR4等从而调节肿瘤细胞生长及转移?
英文摘要:
      Objective:To observe the effect of silencing hypoxia inducible factor-1α(HIF-1α)gene by RNA interference on the proliferation,apoptosis and invasiveness of SGC-7901 cells and to study the molecular mechanism related to these biological changes. Methods:The recombinant plasmid pGCsi-shHIF-1α was transfected into gastric cancer cells SGC-7901 by LipofectamineTM 2000. Then the expressions of HIF-1α in SGC-7901 cells were detected by RT-PCR and Western blot,and the expression of Glut-1,MMP2 and CXCR4 were detected by Western blot. The cell proliferation was detected by MTT assay,and the cell apoptosis was tested by flow cytometry,and the migration and invasiveness of transfected cells were assayed using transwell chambers. Results:After SGC-7901 cells were stably transfected with pGCsi-shHIF-1α,the expression of the mRNA and protein of HIF-1α were inhibited significantly. The expression of Glut-1,MMP-2 and CXCR4 were significantly supressed after HIF-1α silenced by RNA interference. MTT assay showed the proliferation of the pGCsi-shHIF-1α transfected cells was slower than that of empty vector transfected cells. The apoptotic ratio of transfected cells was also higher than that of controls. The migration and invasiveness of the cells transfected with pGCsi-shHIF-1α were significantly suppressed compared with the cells transfected with empty vector. Conclusion:It was showed that the recombinant plasmid pGCsi-shHIF-1α could inhibit the genetic expression of HIF-1α,and inhibit the proliferation,migration and invasiveness,but induce the apoptosis of gastric cancer cells SGC-7901. The results indicated that inhibiting HIF-1α could suppress invasion and migration,and delay tumor growth by down-regulating Glut-1,MMP-2 and CXCR4.
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