Construction of recombinant p16 adenovirus and its suppressive role in the growth of human leukemia cells

First, p16 cDNA was cloned into shuttle plasmid pAdCMV to construct recombinant plasmid pAdCMV-p16, which was cotransfected with pJM17 into the 293 packaging cells by lipofectine-mediated transfection. The prepared recombinant adenovirus was identified by dot blot hybridization and purified by CsCl-gradient ultracentrifuge. The leukemia cell line HL-60, which had a hemizygous deletion and point mutation of p16 gene, was infected with recombinant adenovirus at a multiplicity of infection (M. O. I.) of 50-100 plaqueforming-unit(pfu)/cell. PCR and Western blot indicated that the presence of p16 cDNA and p16 protein was expressed in the infected HL-60; Growth of the Ad-p16 virus-infected HL-60 was inhibited markedly, whereas that of noninfected cells and the cells infected with the control virus Ad-LacZ was not inhibited.