摘要
目的:研究STIM1RNA干扰对人喉癌Hep-2细胞增殖、凋亡、体外侵袭及迁移的影响。方法:采用STIM1干扰慢病毒转染人喉癌Hep-2细胞,实时定量PCR和Western blot验证STIM1干扰效果,CCK-8法分析细胞增殖情况,分别使用Transwell小室侵袭、划痕实验检测细胞体外侵袭和迁移能力,流式细胞术检测细胞凋亡,Western blot分析caspase-3、Bax、Bcl-2、MMP9、VEGF蛋白表达水平。结果:人喉癌Hep-2细胞STIM1干扰慢病毒转染后,干扰组STIM1基因mRNA和蛋白表达均明显减少。与对照组比较,STIM1干扰后,细胞增殖、体外侵袭、迁移能力均明显减弱(均P<0.05);STIM1干扰后Hep-2细胞凋亡率相比对照组显著增大,STIM1干扰促进Hep-2细胞凋亡与上调caspase-3、Bax蛋白表达、下调Bcl-2蛋白表达水平有关;STIM1干扰抑制Hep-2细胞侵袭迁移与下调MMP9、VEGF蛋白表达有关。结论:STIM1RNA干扰可抑制人喉癌Hep-2细胞增殖活性和体外侵袭、迁移能力,且促进喉癌细胞凋亡。
Objective：To study the effects of stromal interaction molecule 1 （STIM1） RNA interference on cell proliferation, apoptosis, invasion and migration ability in vitro in human laryngeal cancer cells Hep-2. Method： Lentivirus transfection was used to realize the STIM1 RNA interference in human laryngeal cancer ceils Hep-2. STIM1 RNA interference was verified by real-time PCR and western blotting. The cell proliferation was analyzed by CCK-8 method, Transwell chamber and wound scratch assay were used to detect cell invasion and migration a- bility, respectively. The cell apoptosis rate was evaluated by flow cytometry. The apoptotic proteins expression levels of caspase-3, Bax, Bcl-2, VEGF and MMP9 were detected by western blotting. Result：The gene expression levels of STIM1 mRNA and protein in human laryngeal cancer cells Hep-2 were significantly decreased after inter- ference lentivirus transfection. After the interference of STIM1, the cell proliferation, invasion and migratory abil ity of Hep-2 cells in vitro declined（P〈0.05）. The cell apoptosis rate significantly increased after STIMI RNA in- terference, along with the caspase-3 and Bax protein expression levels up-regulation, and the Bcl-2 expression lev els down-regulation. The decrease of cell invasion and migratory ability was related with the clown-regulation of VEGF and MMP9 protein expression. Conclusion.. RNA interference of STIMI in human laryngeal cancer cells Hep-2 inhibits the cell proliferation, invasion and migration ability in vitro, and enhances the cell apoptosis rate of Hep-2 cells.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
北大核心
2017年第11期844-848,共5页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
