Regulation of the Mdr1 isoforms in a p53-deficient mouse model

Both p53 and multidrug transporters play important roles in chemoresistance. A transcriptional dependence of the Mdrl gene promoter by p53 was first established a decade ago, and despite intense study, the p53-Mdrl relationship still remains vague in vivo. The general model proposes that wild-type p53 down regulates, while mutant p53 up regulates, the Mdrl promoter. Given that many studies have utilized cancer cell lines, minimal promoters and nonspecific cDNA expression for in vitro experiments, we first sought to confirm the model using dermal fibroblasts isolated from the p53-knockout mice. We show that the gene products of the mouse Mdrl homologue (Mdrla and Mdrlb), namely P-glycoprotein (P-gp), appear upregulated at both the protein and mRNA levels in p53 -/- mFbs compared with p53 +/+ cells. We demonstrate that transient transfection of a mouse p53 WT expression plasmid into short-term primary p53 -/- fibroblasts can revert P-gp overexpression. The difference in P-gp levels has functional significance in that p53 -/- fibroblasts are more resistant to doxorubicin and vincristine treatment and this resistance can be attenuated in the presence of the P-gp inhibitor, verapamil. Furthermore, we demonstrate that in kidney, spleen and testis, P-gp expression is elevated in the absence of p53. In contrast, other organs such as heart, liver, lung, brain, thymus and skeletal muscle, show no difference in expression between p53 +/+ and p53 -/- mice. Thus, our data shows a tissue-specific regulation of P-gp isoforms by p53 in the context of a p53-null mouse model.