Detection of low-level expression of P-glycoprotein in ACHN renal adenocarcinoma cells.

A highly sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) assay and a flow cytometric assay were used to examine ACHN cells for the expression of P-glycoprotein. The expression of P-glycoprotein was detected at the RNA and protein levels in ACHN cells by RT-PCR and flow cytometry, respectively. However, it was below the limit of detection by immunoblotting. The intracellular accumulation of adriamycin in ACHN cells was enhanced by verapamil, cyclosporin A and medroxyprogesterone acetate. Therefore, this study has demonstrated that low-level expression of P-glycoprotein detectable only by RT-PCR and flow cytometry plays a significant role in reducing the intracellular concentration of antitumor agents and thus contributes to the multidrug-resistant phenotype of ACHN cells.