华西口腔医学杂志

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携带细胞周期调控基因p14~(ARF)的重组腺病毒的构建

鲜均明1,周光耀2,梁传余2,刘世喜2   

  1. 1.四川大学基础与法医学院  人体解剖学教研室;2.四川大学华西医院  耳鼻咽喉科,四川  成都 610041
  • 收稿日期:2006-06-25 修回日期:2006-06-25 出版日期:2006-06-20 发布日期:2006-06-20
  • 通讯作者: 刘世喜, Tel: 13982063567
  • 作者简介:鲜均明(1968-), 男, 四川人, 主治医师, 博士
  • 基金资助:

    国家自然科学基金资助项目(0040205401078);四川省青年科技基金资助项目(04ZQ026-024)

Construction of Recombinant Adenovirus Vector Carrying Cell Cycle Controlling Gene-p14ARF

 XIAN Jun-ming1, ZHOU Guang-yao2, LIANG Chuan-yu2, LIU Shi-xi2   

  1. 1. Dept. of Human Anatomy, Institute of Preclinical Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, China; 2. Dept. of Otolaryngology, West China Hospital, Sichuan University, Chengdu 610041, China
  • Received:2006-06-25 Revised:2006-06-25 Online:2006-06-20 Published:2006-06-20

摘要:

目的  构建携带参与细胞周期调控的p14ARF基因的重组腺病毒载体。方法  将野生型p14ARF全长cDNA基因片段插入腺病毒载体质粒pAdTrack-CMV,与骨架质粒在大肠杆菌BJ5183胞内进行同源重组,经293细胞包装、扩增后得到携带p14ARF的重组腺病毒AdEasy-GFP-p14ARF。结果  成功构建了AdEasy-GFP-p14ARF的重组腺病毒载体系统,经测定腺病毒载体滴度达2.3×109 efu/mL。结论  构建的重组腺病毒AdEasy-GFP-p14ARF可望有效地将p14ARF基因导入喉癌细胞株/组织内,为进一步研究喉癌细胞信号转导干预治疗机制提供实验基础。

关键词:  细胞信号转导, 重组腺病毒, 基因治疗, 喉癌

Abstract:

Objective  The recombinant adenovirus vector carrying p14ARF gene was constructed for using in the interference therapy in signal transduction of laryngeal squamous cell carcinoma. Methods  The total cDNA fragment of p14ARF was cloned into the shuttle plasmid pAdTrack-CMV, with the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.Coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was checked by GFP. Results  The recombinant adenovirus vector carrying p14ARF was constructed successfully. The viral titer was 2.3×109. Conclusion  The recombinant adenovirus vector could introduce p14ARF gene into the laryngeal squamous cell carcinoma line or tumor tissue effectively, which would provide experimental basis for the mechanisms and further study of the interference therapy in signal transduction of laryngeal squamous cell carcinoma.

Key words:  cell signal transduction, recombinant adenovirus, gene therapy, laryngeal squamous cell carcinoma