吉林大学学报(医学版)

• 基础研究 • 上一篇    下一篇

siRNA沉默eIF4E诱导人喉癌Hep-2细胞凋亡及其机制

WANG He-bin1,2,WANG Ya-fang1,SHEN Miao-yan2,LI Na3,ZHAO Li-jing2,TENG Bo1   

  1. (1.Department of Otorhinolaryngology and Head-Neck Surgery,Second Hospital,Jilin University,Changchun 130041,China;2. Department of Pathophysiology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China;3. Center of Research and Development,Changchun University of Traditional Chinese  Medicine,Changchun 130117,China)
  • 收稿日期:2013-11-26 出版日期:2014-01-28 发布日期:2014-01-28
  • 通讯作者: 滕 博 E-mail:(Tel:0431-88796796,E-mail:tengbo1975@163.com)
  • 作者简介:王贺彬(1991-),男,河南省周口市人,在读医学硕士,主要从事肿瘤生物治疗研究。
  • 基金资助:

    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)

Apoptosis of human laryngeal  carcinoma  Hep-2 cells induced by siRNA  silencing eIF4E and its mechanism


(1.吉林大学第二医院耳鼻咽喉-头颈外科,吉林 长春 130041;2.吉林大学基础医学院病理生理学系,吉林 长春 130021;3.长春中医药大学研发中心,吉林 长春 130117)     


  1. (1.Department of Otorhinolaryngology and Head-Neck Surgery,Second Hospital,Jilin University,Changchun 130041,China;2. Department of Pathophysiology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China;3. Center of Research and Development,Changchun University of Traditional Chinese  Medicine,Changchun 130117,China)
  • Received:2013-11-26 Online:2014-01-28 Published:2014-01-28

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LipofectamineTM 2000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(P<0.01),Hep-2细胞生存率下降(P<0.05),细胞线粒体膜电位降低(P<0.05),细胞凋亡率增加(P<0.05),凋亡相关基因Bim、Bid及Caspase-3表达上调(P<0.05)。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。

关键词: 小干扰RNA, 真核细胞翻译起始因子4E, 喉肿瘤, Hep-2细胞, 细胞凋亡

Abstract:

To observe the effect of the eukaryotic translation initiation factor 4E (eIF4E) targeted small interfering RNA (siRNA) on the eIF4E expression in human laryngeal Hep-2 cells,and to clarify the induction effect on apoptosis of Hep-2 cells and its mechanism.Methods The siRNA-eIF4E was transfected into human laryngeal carcinoma Hep-2 cells (siRNA-eIF4E group) with LipofectamineTM 2000,at the same time blank control group and empty plasmid group were set up.The inhibitory rate of cell proliferation in each group was detected by MTT.The changes of mitochondrial membrane potentials of the cells were observed by Rhodamine dyes.Apoptosis was detected by TUNUL staining,and the transcription and protein expression levels  of apoptosis related genes were analyzed by RT-PCR and Western blotting method.Results Compared with blank control group and empty plasmid group,the transcription and protein expression levels of eIF4E in Hep-2 cells in siRNA-eIF4E group were significantly decreased(P<0.01),the survival rate of Hep-2 cells was  decreased (P<0.05),the mitochondrial membrane potential of the cells was decreased (P<0.05),the apoptotic rate was increased (P<0.05),and the expression levels of apoptosis related genes Bim,Bid and Caspase3 were significantly up-regulated (P<0.05).Conclusion siRNA-eIF4E can inhibit the proliferation of human laryngeal carcinoma  Hep-2 cells in vitro,and its mechanism may be related to the activation of mitochondrial apoptosis pathway.

Key words: small interfering RNA, eukaryotic translation initiation factor 4E, laryngeal neoplasms, Hep-2 cells, apoptosis

中图分类号: 

  • R739.65