siRNA沉默eIF4E诱导人喉癌Hep-2细胞凋亡及其机制

发布时间：2018-02-27 13:12

本文关键词： 小干扰RNA 真核细胞翻译起始因子E 喉肿瘤 Hep-细胞细胞凋亡　出处：《吉林大学学报(医学版)》2014年01期 　论文类型：期刊论文

【摘要】：目的:观察真核细胞翻译起始因子4E(eIF4E)基因的靶向小干扰RNA(siRNA)对喉癌Hep-2细胞中eIF4E基因表达的影响,探讨其诱导喉癌细胞凋亡的作用机制。方法:采用脂质体LipofectamineTM2000将siRNA-eIF4E转染入人喉癌Hep-2细胞(siRNA-eIF4E组),同时设空白对照组和空质粒组。MTT法检测siRNA对Hep-2细胞增殖的抑制作用,罗丹明染色检测转染后细胞内线粒体膜电位的变化,TUNEL染色法检测细胞凋亡,RT-PCR和Western blotting法检测凋亡相关基因转录和蛋白表达水平的变化。结果:与空白对照组及空质粒组比较,siRNA-eIF4E组Hep-2细胞中eIF4E基因转录和表达水平明显下调(P0.01),Hep-2细胞生存率下降(P0.05),细胞线粒体膜电位降低(P0.05),细胞凋亡率增加(P0.05),凋亡相关基因Bim、Bid及Caspase-3表达上调(P0.05)。结论:siRNA-eIF4E在体外可抑制人喉癌Hep-2细胞增殖,其机制可能是通过激活线粒体凋亡途径诱导Hep-2细胞凋亡。
[Abstract]:Aim: to observe the effect of targeting small interfering RNAs (siRNAs) of eukaryotic cell translation initiation factor 4EIF4E on the expression of eIF4E gene in Hep-2 cells of laryngeal carcinoma. Methods: siRNA-eIF4E was transfected into human laryngeal carcinoma Hep-2 cells by lipofectin LipofectamineTM2000 into siRNA-eIF4E group. The inhibitory effect of siRNA on the proliferation of Hep-2 cells was detected by blank control group and empty plasmid group. The changes of mitochondrial membrane potential after transfection were detected by Rhodamine staining and the expression of apoptosis-related genes by RT-PCR and Western blotting were detected by Tunel staining. Results: compared with the blank control group and empty substance, the changes of apoptosis-related gene transcription and protein expression were detected by Tunel staining. The transcription and expression of eIF4E gene in Hep-2 cells were significantly down-regulated by siRNA-eIF4E group. The survival rate of Hep-2 cells was decreased, the mitochondrial membrane potential was decreased, the apoptosis rate was increased, and the expression of the apoptosis-related gene BimsiRNA-eIF4E was increased. Conclusion\% siRNA-eIF4E can increase the expression of BimsiRNA-eIF4E in Hep 2 cells. It can inhibit the proliferation of human laryngeal carcinoma Hep-2 cells in vitro. The mechanism may be the activation of mitochondrial apoptosis pathway to induce apoptosis of Hep-2 cells.
【作者单位】：吉林大学第二医院耳鼻咽喉-头颈外科;吉林大学基础医学院病理生理学系;长春中医药大学研发中心;
【基金】：国家自然科学基金资助课题(81302206) 吉林省科技厅自然科学基金资助课题(201105106)
【分类号】：R739.65

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