Mutant p53 R248Q but not R248W enhances in vitro invasiveness of human lung cancer NCI-H1299 cells

Abstract

More than half of all human cancers are associated with mutations of the TP53 gene. In regard to the functional interaction with the remaining wild-type (WT) p53 allele, p53 mutations are classic-fed into two types, recessive and dominant-negative (DN) mutations. The latter mutant protein has a DN activity over the remaining WT allele. We previously showed that the DN p53 mutant was useful as a predictor of poor outcome or a risk factor for meta static recurrence in patients with some types of cancers, regardless of the presence or absence of loss of heterozygosity (LOH) of WT p53, suggesting that the DN p53 had 'gain-of-function (GOF)' activity besides the trans-dominance function. In this study, we investigated GOF activity of two DN p53 mutants which had a point mutation at cordon 248 (R248Q and R248W), one of the hot spots, by transecting them respectively into H1299 cells which originally expressed no p53 protein. Growth activity of the transfect ants with the two mutants was not different from that of parent or Mock transfect ants. Meanwhile, in vitro invasions of Materiel and type I collagen gel by R248Q-transfectants were significantly higher than those by R248W-transfectants or the control cells. However, there were no differences in cell motile activities, expressions of extra cellular matrix-derivative enzymes such as matrix metalloproteinases, urokinase-type plasminogen activator and heparins, and their inhibitors, between R248Q- and R248W-transfectants. These findings indicate that the p53 mutants have a different quality in GOF activities even if the mutations occurred at the same cordon. And detailed information of the status of p53, including Tran dominancy and GOF activity, is expected to be useful for diagnosis and therapeutic strategy fitting the individual patients.