Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry

N. Feller, C. M. Kuiper, J. Lankelma, J. K. Ruhdal, R. J. Scheper, H. M. Pinedo, H. J. Broxterman

Research output: Contribution to journalArticle

Abstract

Multidrug resistance (MDR) in tumour cells is often caused by the overexpression of the plasma membrane drug transporter P-glycoprotein (P-gp) or the recently discovered multidrug resistance-associated protein (MRP). In this study we investigated the specificity and sensitivity of the fluorescent probes rhodamine 123 (R123), daunorubicin (DNR) and calcein acetoxymethyl ester (calcein-AM) in order to detect the function of the drug transporters P-gp and MRP, using flow cytometry. The effects of modulators on the accumulation and retention of these probes were compared in several pairs of sensitive and P-gp- as well as MRP-overexpressing cell lines. R123, in combination with the modulator PSC833, provided the most sensitive test for detecting P-gp-mediated resistance. Moreover, in a 60 min drug accumulation assay R123 can be regarded as a P-gp-specific probe, since R123 is not very efficiently effluxed by MRP. In contrast to R123, a 60 min DNR or calcein-AM accumulation test could be used to detect MRP-mediated resistance. The MRP-specific modulator genistein could be used in combination with DNR, but not with calcein-AM. Vincristine (VCR) can be used to increase the cellular uptake of calcein-AM in MDR cells, but is not specific for MRP. Thus, although the combination of DNR with genistein appeared to be as sensitive as the combination of calcein-AM with VCR, the former may be used to probe specific MRP activity whereas the latter provides a combined (P-gp + MRP) functional MDR parameter. With these functional assays the role and relative importance of P-gp and MRP can be studied in, for example, haematological malignancies.

Original languageEnglish (US)
Pages (from-to)543-549
Number of pages7
JournalBritish Journal of Cancer
Volume72
Issue number3
StatePublished - 1995
Externally publishedYes

Fingerprint

Multidrug Resistance-Associated Proteins
Multiple Drug Resistance
Flow Cytometry
P-Glycoprotein
Rhodamine 123
Daunorubicin
Neoplasms
Genistein
Vincristine
Pharmaceutical Preparations
Membrane Transport Proteins
Hematologic Neoplasms
Fluorescent Dyes
Cell Membrane
calcein AM

Keywords

  • Calcein
  • Daunorubicin
  • Multidrug resistance-associated protein
  • P-glycoprotein
  • PSC833
  • Rhodamine

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Feller, N., Kuiper, C. M., Lankelma, J., Ruhdal, J. K., Scheper, R. J., Pinedo, H. M., & Broxterman, H. J. (1995). Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry. British Journal of Cancer, 72(3), 543-549.

Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry. / Feller, N.; Kuiper, C. M.; Lankelma, J.; Ruhdal, J. K.; Scheper, R. J.; Pinedo, H. M.; Broxterman, H. J.

In: British Journal of Cancer, Vol. 72, No. 3, 1995, p. 543-549.

Research output: Contribution to journalArticle

Feller, N, Kuiper, CM, Lankelma, J, Ruhdal, JK, Scheper, RJ, Pinedo, HM & Broxterman, HJ 1995, 'Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry', British Journal of Cancer, vol. 72, no. 3, pp. 543-549.
Feller N, Kuiper CM, Lankelma J, Ruhdal JK, Scheper RJ, Pinedo HM et al. Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry. British Journal of Cancer. 1995;72(3):543-549.
Feller, N. ; Kuiper, C. M. ; Lankelma, J. ; Ruhdal, J. K. ; Scheper, R. J. ; Pinedo, H. M. ; Broxterman, H. J. / Functional detection of MDR1/P170 and MRP/P190-mediated multidrug resistance in tumour cells by flow cytometry. In: British Journal of Cancer. 1995 ; Vol. 72, No. 3. pp. 543-549.
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