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.
Kawamoto
S
S
Department of Urology, Gifu University School of Medicine, Japan.
Deguchi
T
T
Nezasa
S
S
Yamada
S
S
Okano
M
M
Kawada
Y
Y
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000032:analysis*; D000903:Antibiotics, Antineoplastic / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000737:chemistry*; D004317:Doxorubicin / Q000378:metabolism; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D006801:Humans; D007680:Kidney Neoplasms / Q000737:chemistry*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D012194:RNA-Directed DNA Polymerase; D014407:Tumor Cells, Cultured
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000903:Antibiotics, Antineoplastic; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D004317:Doxorubicin; D012194:RNA-Directed DNA Polymerase