PMID: 7622426Jun 1, 1995Paper

New flow cytometric method for detection of minimally expressed multidrug resistance P-glycoprotein on normal and acute leukemia cells using biotinylated MRK16 and streptavidin-RED670 conjugate

Japanese Journal of Cancer Research : Gann
Akihiro TakeshitaR Ohno

Abstract

To evaluate the expression of multidrug resistance (MDR) on normal and leukemia cells, we examined P-glycoprotein (P-gp) by a newly devised flow cytometric method, utilizing a biotinylated monoclonal antibody (mAb) against P-gp (MRK16), a streptavidin-RED670 conjugate (SA-RED670) and appropriate emission filters. The combination of biotinylated MRK16 (b-MRK16) and SA-RED670 resulted in higher sensitivity as compared with standard methods such as the use of streptavidin-phycoerythrin (SA-PE) conjugate. The sensitivity was examined in K562, K562/ADR, NOMO-1, NOMO-1/ADR and HL60 cells, and compared with the data obtained from reverse transcription polymerase chain reaction (RT-PCR) of mdr-1 gene. P-gp positivity on flow cytometry was 10.4%, 99.9%, 1.4%, 90.4% and 0%, respectively. Mdr-1 mRNA was well expressed in K562/ADR and NOMO-1/ADR cells, but not in NOMO-1 and HL60 cells. In K562 cells, mdr-1 was found after 40 cycles of PCR, but not 25 cycles. These data are well correlated with those from the flow cytometry. We then studied the P-gp expression on normal peripheral blood cells and acute leukemia cells. P-gp was little expressed on peripheral lymphocytes, monocytes and granulocytes. It was also little expressed on blast cells...Continue Reading

References

Sep 1, 1992·Leukemia & Lymphoma·A F List, C M Spier
Sep 1, 1992·British Journal of Haematology·C LudescherJ Hofmann
Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·E GeorgesV Ling
May 15, 1991·Journal of the National Cancer Institute·R PirkerL Havelec
Oct 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·H Hamada, T Tsuruo
Oct 1, 1989·Japanese Journal of Cancer Research : Gann·Y IshidaM Shimoyama

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Citations

Aug 27, 2003·Journal of Tongji Medical University = Tong Ji Yi Ke Da Xue Xue Bao·Z ChenR Ohno
May 3, 2008·Biological & Pharmaceutical Bulletin·Kayo HenmiToshihiko Hirano

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