Expression of TRAIL mediated by survivin promoter in cervical cancer Hela cell

Objective: To explore the potential use of TRAIL expression vector mediated by survivin promoter for the targeted gene therapy in cervical cancer.Methods: The total RNA was extracted from human peripheral blood lymphocytes(PBMCs) and was used as Trail PCR template,TRAIL gene with interleukin 2(IL-2) signal peptide gene was amplified by RT-PCR method using primers with IL-2 gene sequence in 5' end of primer,and the PCR products were cloned into the vector pGL-Basic/surp downstream survivin promoter to form an eukaryotic expression vector that was then transfected into Hela and normal hepatocyte(7702).Expression of TRAIL was identified by RT-PCR and western blot from transfected cell.Results: A 613 bp cDNA fragment was amplified from PBMCs RNA and was further conformed by sequencing as IL-2 and trail fusion gene which was cloned into pGL-Basic/surp vector downstream survivin promoter to form eukaryotic expression vector pGL-Basic/surp/TRAIL.Enzyme digestion showed the correct recombinant.The results of the RT-PCR and western blot showed that pGL-Basic/surp/TRAIL vector tranfected Hela cells successfully expressed TRAIL,while there was no TRAIL expression in 7702 cell.Conclusion: The successful construction of survivin promoter controlling TRAIL eukaryotic expression vector provides the possibility for effective gene therapy of cervical cancer.