Verapamil enhances doxorubicin activity in cultured human renal carcinoma cells

Abstract

Cells from 22 renal cell carcinomas (RCC) were established in culture. Sensitivity of the tumour cells to doxorubicin alone and in combination with racemic verapamil, which reverses multidrug resistance, was tested using a [ 75Se]selenomethionine uptake assay to measure protein synthesis. The effect of verapamil was expressed as a potentiation index: ld 50doxorubicin/ ld 50doxorubicin + verapamil. The potentiation index in 15 of these carcinomas was determined for cells within the first 14 days of culture. At 3.3 μmol/l concentration of verapamil, of the tumours sensitive to doxorubicin alone ( ld 50 < 0.75 μg/ml) five of seven showed a potentiation index of > 2. For the less sensitive tumours the analogous proportion was seven of eight. Tumour cell expression of glycoprotein P-170, associated with multidrug resistance, was estimated using the monoclonal antibody C-219. Initial expression levels were unrelated to the action of verapamil. In five tumours the proportion of cells expressing P-170 declined as the period of culture increased. This was not associated with any consistent change in the ld 50 for doxorubicin or in potentiation of doxorubicin sensitivity by verapamil. Cell cloning associated with prolonged cell growth in vitro could mimic tumour cell cloning which accompanies the formation of metastases. Thus reduced expression of P-170 on prolonged cell growth in vitro may be a pointer to the efficacy of combination therapy in the treatment of patients with metastatic renal cell carcinoma.