Enhancement of sensitivity of urinary bladder tumor cells to cisplatin by c-myc antisense oligonucleotide

Abstract

Tumor cells have different degrees of sensitivity and resistance to anticancer agents. The acquisition of drug resistance is a major concern in cancer treatment. Because the c-myc oncogene has been implicated in the poor prognosis of some human tumors including urinary bladder tumor (UBT), the resistance of such tumors to anticancer drugs may correlate with c-myc expression. The present study tested this hypothesis by examining the effect of c-myc antisense oligonucleotide treatment on the sensitivity of human UBT cells to anticancer chemotherapeutic agents. Cytotoxicity determined by a 1-day microculture tetrazolium dye assay. C-myc mRNA was examined by Northern blot analysis. The T24 human UBT cell line constitutively expresses c-myc mRNA. Treatment of T24 cells with the c-myc antisense oligonucleotide resulted in a significant inhibition of the expression of c-myc mRNA. Treatment of T24 cells with the c-myc antisense oligonucleotide in combination with mitomycin C, Adriamycin, or 5-fluorouracil (5-FU) did not overcome their resistance to these anticancer chemotherapeutic agents. However, combination treatment with the c-myc antisense oligonucleotide and CDDP resulted in a synergistic cytotoxic effect on T24 cells and two freshly derived UBT cells. Further, treatment of CDDP-resistant T24 cells (T24/CDDP) with c-myc antisense oligonucleotide and CDDP reversed the resistance. Pretreatment of T24 cells with either agent and then treatment with the second agent resulted in the same cytotoxic activity as achieved in the presence of the two agents. The combination of c-myc antisense oligonucleotide and carboplatin also resulted in a synergistic cytotoxic effect on T24 cells, and the combination of c-myc antisense oligonucleotide and trans-diamminedichloro-platinum resulted in an additive cytotoxic effect. Incubation of T24 or T24/CDDP cells with the c-myc antisense oligonucleotide increased the intracellular accumulation of CDDP, but not the accumulation of 5-FU. This study demonstrates that combination treatment with c-myc antisense oligonucleotide and CDDP can overcome the CDDP-resistance of UBT cells and that the increased intracellular accumulation of CDDP by c-myc antisense oligonucleotide may play a role in the enhanced cytotoxicity obtained. The synergistic effect obtained with established UBT cells and freshly isolated UBT cells suggests that combination treatment with c-myc antisense oligonucleotide and CDDP may have clinical application in the therapy of CDDP-resistant UBT.