title: Gain-of-Function Activity of Mutant p53 in Lung Cancer through Up-Regulation of Receptor Protein Tyrosine Kinase Axl.
abstract: p53 mutations are present in up to 70% of lung cancer. Cancer cells with p53 mutations, in general, grow more aggressively than those with wild-type p53 or no p53. Expression of tumor-derived mutant p53 in cells leads to up-regulated expression of genes that may affect cell growth and oncogenesis. In our study of this aggressive phenotype, we have investigated the receptor protein tyrosine kinase Axl, which is up-regulated by p53 mutants at both RNA and protein levels in H1299 lung cancer cells expressing mutants p53-R175H, -R273H, and -D281G. Knockdown of endogenous mutant p53 levels in human lung cancer cells H1048 (p53-R273C) and H1437 (p53-R267P) led to a reduction in the level of Axl as well. This effect on Axl expression is refractory to the mutations at positions 22 and 23 of p53, suggesting that p53's transactivation domain may not play a critical role in the up-regulation of Axl gene expression. Chromatin immunoprecipitation (ChIP) assays carried out with acetylated histone antibodies demonstrated induced histone acetylation on the Axl promoter region by mutant p53. Direct mutant p53 nucleation on the Axl promoter was demonstrated by ChIP assays using antibodies against p53. The Axl promoter has a p53/p63 binding site, which however is not required for mutant p53-mediated transactivation. Knockdown of Axl by Axl-specific RNAi caused a reduction of gain-of-function (GOF) activities, reducing the cell growth rate and motility rate in lung cancer cells expressing mutant p53. This indicates that for lung cancer cell lines with mutant p53, GOF activities are mediated in part through Axl.
authors.affiliation: Department of Biochemistry & Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Vaughan
authors.forename: Shilpa
authors.initials: S
authors.lastname: Singh
authors.forename: Brad
authors.initials: B
authors.lastname: Windle
authors.forename: W Andrew
authors.initials: WA
authors.lastname: Yeudall
authors.forename: Rebecca
authors.initials: R
authors.lastname: Frum
authors.forename: Steven R
authors.initials: SR
authors.lastname: Grossman
authors.forename: Swati P
authors.initials: SP
authors.lastname: Deb
authors.forename: Sumitra
authors.initials: S
authors.lastname: Deb