Comparison of (99m)Tc-sestamibi and doxorubicin to monitor inhibition of P-glycoprotein function.
P-glycoprotein (Pgp) overexpression is a well-recognized factor in resistance to chemotherapy. Doxorubicin flow cytometry is used to monitor Pgp function in haematological specimens and biopsies from other cancers, and radionuclide imaging with sestamibi has recently shown promise for non-invasive monitoring. In the present study the two methods were directly compared in single-cell suspensions of three variants of the human breast carcinoma cell line MCF7: sensitive MCF7/WT, doxorubicin-selected MCF7/AdrR, and MDR1-gene-transfected MCF7/BC19 cells with doxorubicin resistance factors of 1, 192, and 14, respectively. Accumulation of sestamibi and mean fluorescence of doxorubicin (5.5 microM) were assessed over 60 min in the presence and absence of Pgp modulators GG918 (0.01 to 0.2 microM) and PSC833 (0.05 to 2.0 microM). Accumulation curves for sestamibi and doxorubicin differed among the cell variants under control conditions, with sestamibi showing a significantly greater difference between WT and resistant cells than doxorubicin. Both GG918 and PSC833 reversed uptake deficits to WT levels for sestamibi in MCF7/BC19 cells and doxorubicin in MCF7/BC19 and MCF7/AdrR cells, but failed to show the same effect for sestamibi in MCF7/AdrR cells (approximately 30% of MCF7/WT level). Thus, both methods clearly distinguished sensitive from resistant MCF7 variants, with the radionuclide method showing greater sensitivity.
Muzzammil
T
T
Department of Pharmaceutical Sciences, University of Toronto, 610 University Avenue, Toronto, Ontario, M5G 2M9, Canada.
Moore
M J
MJ
Hedley
D
D
Ballinger
J R
JR
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D000166:Acridines / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000493:pharmacokinetics* / Q000494:pharmacology; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D003524:Cyclosporins / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000493:pharmacokinetics* / Q000494:pharmacology; D019450:Genes, MDR / Q000235:genetics / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D007546:Isoquinolines / Q000494:pharmacology; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics*; D017256:Technetium Tc 99m Sestamibi / Q000493:pharmacokinetics*; D044005:Tetrahydroisoquinolines; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D018435:ATP Binding Cassette Transporter, Subfamily B; D000166:Acridines; D000970:Antineoplastic Agents; D003524:Cyclosporins; D007546:Isoquinolines; D019275:Radiopharmaceuticals; D044005:Tetrahydroisoquinolines; D004317:Doxorubicin; D017256:Technetium Tc 99m Sestamibi; C083501:Elacridar; C070272:valspodar