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

作　　者：曾楷峰[1] 施瑞华[1] 朱宏[1] 张国新[1] 郝波[1] 

机构地区：[1]南京医科大学第一附属医院消化科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2010年第2期144-149,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助项目(30770991)

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。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 as...更多say showed the proliferation of the pGCsi-shHIF-1α transfected cells was slower than that of empty vector transfected cells. The apoptotic ra tio 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.

关 键 词：胃癌 缺氧诱导因子-1a RNA干扰 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]