Construction of recombinant adenovirus vector of Livin siRNA and its effects on proliferation,apoptosis and drug resistance of K562 cells

Objective To study the effect of recombinant adenovirus vector-mediated Livin siRNA on the proliferation,apoptosis and drug resistance in K562 cells.Methods The targeting Livin siRNA was subcloned into the shuttle plasmid pSES-HUS,and then recombinant it with adenovirus skeleton plasmid pAdeasy by homologous recombination.We transfected the product into HEK293 cells via liposome and generated recombinant adenovirus Ad5-Livin siRNA.K562 cells were infected with Ad5-Livin siRNA.The gene silencing efficiency of recombinant adenovirus Ad5-Livin siRNA was monitored by real-time PCR and Western blot analysis in K562 cells.The relationship between Livin siRNA and K562 cells was detected by MTT assays and flow cytometry.Results The recombinant adenovius vector of Ad5-Livin siRNA was successfully constructed and infected into K562 cells.Recombinant adenovirus vector silenced Livin gene at the level of mRNA and protein,inhibited proliferation and enhanced apoptosis and sensitivity to Adriamycin,Vincristine and Etoposide in K562 cells.Conclusion The recombinant adenovirus Ad5-Livin siRNA effectively silences the expression of Livin gene and enhances the apoptosis and sensitivity to anticancer drugs in K562 cells.Livin is a new molecular target of leukemia's therapy.