Recombinant adenovirus expressing Von Hippel-Lindau-mediated cell cycle arrest is associated with the induction of cyclin-dependent kinase inhibitor p27Kip1

A recombinant adenovirus containing the Von Hippel-Lindau (VHL) cDNA was constructed (AdVHL) and used to investigate the function of this tumor suppressor gene. Exposure of renal and breast cancer cell lines to AdVHL resulted in high levels of VHL mRNA and protein. AdVHL infection resulted in G1 cell cycle arrest and growth inhibition of renal and breast cancer cell lines. AdVHL-mediated cell cycle arrest was associated with induction of the cyclin-dependent kinase inhibitor (CDKI) p27(Kip1) and inhibition of CDK2 and cyclinB1-dependent cdc2 activities. Nuclear run-on analyses and actinomycin D inhibition studies indicate that the induction of p27(Kip1) RNA by VHL is mediated at least in part through an increase in p27(Kip1) mRNA synthesis. Furthermore, [S-35]methionine pulse-chase studies indicate that the increase in p27(Kip) expression is also regulated through posttranscriptional control mechanisms. These studies support a novel concept that the tumor suppressor gene VHL controls cell cycle progression by regulation of p27(Kip1) at both the mRNA and protein levels. (C) 1998 Academic Press.