Modulation of cellular chemoresistance in keratinocytes by activation of different oncogenes.

Abstract

Response to chemotherapeutic agents in malignant tumors depends on many factors, most of which are as yet unknown. We investigated the correlation between the activation of different oncogenes and protein-kinase-C (PKC) modulation, and the cytotoxicity of some of the most widely used anti-cancer drugs. We transformed the murine keratinocyte cell line PAM 212, with different oncogenes (v-H-ras, v-myc and adenovirus E1a) and a mutant p53 suppressor gene (mp53). The cytotoxic effect of cisplatin (CDDP), doxorubicin (DOX) and vincristine (VCR), together with the concomitant action of modulators of PKC, TPA and staurosporine were evaluated by the crystal-violet method, thymidine incorporation and flow cytometry. We report that (a) the oncogene v-H-ras induces resistance to CDDP (> 50%), DOX (> 25%) and VCR (> 20%); (b) the E1a oncogene induces only resistance to VCR (> 40%) and marked sensitivity to CDDP and DOX; (c) the mp53 oncogene induces more resistance to VCR and insignificant resistance to the other drugs; and (d) activation of PKC by TPA increases the resistance to VCR and DOX in cells transformed by the v-H-ras, while it significantly increases the lethality with CDDP of the E1a-transformed cells. Staurosporine increases the cytoxicity of all the drugs, especially in the E1a-transformed keratinocytes. In the flow-cytometry analysis, the percentage of BUdR incorporation was related to sensitivity to anti-cancer drugs.