Construction of eukaryotic co expression plasmid pBud-TRAIL-IL-24 and its expression in cervical cancer cells

Abstract:

Objective:To construct the eukaryotic co expression plasmid encoding TNF related apoptosis inducing ligand(TRAIL)gene and interleukin 24(IL-24)gene and to study its inhibitory effect on cervical cancer cell lines.Methods:IL-24 gene was inserted into the expression vector of pBud-TRAIL to obtain co-expression plasmid pBud-TRAIL-IL-24 by molecular cloning technology.The constructed co-expression plasmid was identified by restriction endoenzyme digestion and nucleotides sequencing.The recombinant plasmid of pBud-TRAIL-IL-24 was transfected into HeLa cells.The expressions of TRAIL and IL-24 were detected by fluorescence microscopy and immunofluorescence double labeling staining.The proliferation of cervical cancer cells was assessed by MTT assay．Results:The eukaryotic co-expression plasmid pBud-TRAIL-IL-24 could be successfully constructed and be efficiently co-expressed in HeLa cells.The recombinant plasmid significantly inhibited the growth of cervical cancer cells(P<0.05).Conclusion:The plasmid pBud-TRAIL-IL-24 can be successfully constructed and efficiently co-expressed in HeLa cells,and it can effectively inhibit the proliferation of HeLa cells,which lays the foundation for further study of the joint application of TRAIL and IL-24 in cervical cancer gene therapy.