含hIL-24重组腺病毒载体(Ad-mda-7/hIL-24)的构建及其对喉癌细胞Hep2增殖的影响  

The construction of recombinant adenovirus vector containing hIL-24 and its effect on the Hep2 laryngeal carcinoma cells

作　　者：刘娣[1] 许晓群[1] 张建华[1] 陈雪梅[2] 苏庆红[1] 孟德萍[1] 王郡甫[1] 

机构地区：[1]山东省医学科学院基础医学研究所山东省现代医用药物与技术重点实验室,山东济南250062 [2]山东大学第二医院,山东济南250033

出　　处：《中国生化药物杂志》2012年第2期121-125,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：山东省中青年科学家奖励基金(BS2009SW007);山东省自然科学基金(ZR2009CM036和ZR2010CM067)

摘　　要：目的构建含有人白细胞介素24(hIL-24)的腺病毒表达载体(Ad-mda-7/hIL-24),观察其对喉癌细胞Hep2增殖的影响。方法提取IL-24cDNA克隆至穿梭质粒pAdTrack-CMV,采用pAdEasy系统进行细菌内同源重组得到含目的基因的腺病毒载体,脂质体转染293A细胞进行包装并扩增病毒。PCR和Western blot方法鉴定重组腺病毒,四甲基偶氮唑盐(MTT)法观察其对Hep2增殖的影响。结果成功构建Ad-mda-7/hIL-24,滴度达107pfu/mL,MTT检测表明Ad-mda-7/hIL-24可明显抑制Hep2细胞生长。结论构建有较强感染能力的Ad-mda-7/hIL-24,并能显著抑制Hep2细胞增殖,为研究其作用机制及将其用于喉癌的基因治疗提供了实验和理论依据。Purpose To construct recombinant adenovirus vector containing hIL-24 gene（Ad-mda-7/hIL-24） and to detect its inhibition on the proliferation of Hep2 laryngeal carcinoma cells.MethodsThe hIL-24 cDNA was cloned into the shuttle plasmid pAdTrack-CMV.The Ad-mda-7/hIL-24 was constructed by the method of homogenous recombination in bacteria BJ5183 containing the PAdEasy system.The correct recombinant was packaged with liposome and transfected into 293A cells to construct and amplify virus.PCR and Western blot method were used to identify the recombinant adenovirus,tetrazolium salt（MTT） method to observe the effect on proliferation of Hep2.ResultsRestriction endonuclease and PCR analysis confirmed that the hIL-24 gene was successfully inserted into the adenovirus vector.The titer of the recombinant adenovirus was 107 pfu/mL.MTT assay showed that Ad-mda-7/IL-24 significantly inhibited the proliferation of Hep2 cells.ConclusionThe recombinant adenovirus vector-hIL-24 was successfully constructed,which laid a foundation for studying Ad-mda-7/IL-24 gene in gene therapy of laryngeal cancer.

关 键 词：hIL-24 腺病毒 癌 喉 细胞增殖 

分 类 号：Q78[生物学—分子生物学] R966[医药卫生—药理学]