Construction recombinant adenovirus vector expressing human interleukin-24 and analysis of its biological activity in vitro

Objective:To clone human IL-24 gene,construct its recombinant adenovirus vector,and to study its biological activity.Methods:The expression of IL-24 in HeLa cells was induced by mezerein,IL-24 cDNA was amplified by RT-PCR and then was subcloned to the vector of pAdTrack-CMV.Generation of recombinants by cotransforming the PmeⅠ-cut shuttle plasmids with pAdEasy-1 backbone vectors in BJ5183,virus was multiplied in 293 cells and was identified by PCR and Western blot.The inhibitory effect of IL-24 to CaSki cells was detected by MTT assay and cell survival analysis.Apoptosis was detected by Hoechst 33342 stain and flow cytometry.The expression of Bax,Bcl-2 and p53 was detected by Western blot.Results:The sequence of IL-24 gene was identical with that in GeneBank,the recombinant adenovirus vector was successfully constructed,this study showed that AdIL-24 induced growth suppression and apoptosis of CaSki cells.The Bax and p53 protein were up regulated while Bcl-2 protein was down regulated.Conclusion:The adenovirus vector of hIL-24 was successfully constructed and adenovirus recombinants had biological activity.