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

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.