Quantitative analysis of breast cancer resistance protein and cellular resistance to flavopiridol in acute leukemia patients

Takeo Nakanishi, Judith Karp, Ming Tan, L. Austin Doyle, Todd Peters, Weidong Yang, David Wei, Douglas D. Ross

Research output: Contribution to journalArticle

Abstract

Purpose: Flavopiridol is a cyclin-dependent kinase inhibitor currently undergoing human clinical trials. As clinical development is pursued, it becomes important to evaluate resistance mechanisms to flavopiridol. To elucidate the contribution of breast cancer resistance protein (BCRP) to cellular resistance to flavopiridol in acute myeloid leukemia, we studied the relationship between cellular resistance to flavopiridol and mRNA expression of BCRP or P-glycoprotein (P-gp, product of MDR1 gene) in blast cells from adult patients with acute leukemia. Experimental Design: Twenty-one blast cell samples from 20 patients were studied. The expression of BCRP, P-gp, β-actin mRNA was determined by real-time reverse transcription-PCR, using fluorescent hybridization probes to evaluate codon 482, a known site of mutations in BCRP mRNA. In vitro cell viability and apoptosis were examined after 24 h exposure to flavopiridol. Results: BCRP mRNA expression varied over a 200-fold range. In the blast cell samples with BCRP mRNA expression > 10,000 copies/pg β-actin (n = 9), BCRP mRNA correlated proportionally with cell viability in the presence of 250 nM flavopiridol (r = 0.86, P = 0.003) and with apoptosis induced by flavopiridol (r = 0.71, P = 0.031). In contrast, MDR1 mRNA expression did not correlate with either flavopiridol cytotoxicity or induction of apoptosis. Melting point analysis of the hybridization probes determined that all 21 patient samples had arginine at codon 482 of BCRP mRNA, the wild-type form. Conclusions: These results suggest that unlike P-gp, BCRP may play a role in leukemia cellular resistance to flavopiridol. No mutations at codon 482 were observed in BCRP mRNA in this group of patients.

Original languageEnglish (US)
Pages (from-to)3320-3328
Number of pages9
JournalClinical Cancer Research
Volume9
Issue number9
StatePublished - Sep 1 2003
Externally publishedYes

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alvocidib
Leukemia
Breast Neoplasms
Messenger RNA
Proteins
Codon
Apoptosis
Actins
Cell Survival
Mutation
Cyclin-Dependent Kinases

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Nakanishi, T., Karp, J., Tan, M., Austin Doyle, L., Peters, T., Yang, W., ... Ross, D. D. (2003). Quantitative analysis of breast cancer resistance protein and cellular resistance to flavopiridol in acute leukemia patients. Clinical Cancer Research, 9(9), 3320-3328.

Quantitative analysis of breast cancer resistance protein and cellular resistance to flavopiridol in acute leukemia patients. / Nakanishi, Takeo; Karp, Judith; Tan, Ming; Austin Doyle, L.; Peters, Todd; Yang, Weidong; Wei, David; Ross, Douglas D.

In: Clinical Cancer Research, Vol. 9, No. 9, 01.09.2003, p. 3320-3328.

Research output: Contribution to journalArticle

Nakanishi, T, Karp, J, Tan, M, Austin Doyle, L, Peters, T, Yang, W, Wei, D & Ross, DD 2003, 'Quantitative analysis of breast cancer resistance protein and cellular resistance to flavopiridol in acute leukemia patients', Clinical Cancer Research, vol. 9, no. 9, pp. 3320-3328.
Nakanishi, Takeo ; Karp, Judith ; Tan, Ming ; Austin Doyle, L. ; Peters, Todd ; Yang, Weidong ; Wei, David ; Ross, Douglas D. / Quantitative analysis of breast cancer resistance protein and cellular resistance to flavopiridol in acute leukemia patients. In: Clinical Cancer Research. 2003 ; Vol. 9, No. 9. pp. 3320-3328.
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AU - Yang, Weidong

AU - Wei, David

AU - Ross, Douglas D.

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N2 - Purpose: Flavopiridol is a cyclin-dependent kinase inhibitor currently undergoing human clinical trials. As clinical development is pursued, it becomes important to evaluate resistance mechanisms to flavopiridol. To elucidate the contribution of breast cancer resistance protein (BCRP) to cellular resistance to flavopiridol in acute myeloid leukemia, we studied the relationship between cellular resistance to flavopiridol and mRNA expression of BCRP or P-glycoprotein (P-gp, product of MDR1 gene) in blast cells from adult patients with acute leukemia. Experimental Design: Twenty-one blast cell samples from 20 patients were studied. The expression of BCRP, P-gp, β-actin mRNA was determined by real-time reverse transcription-PCR, using fluorescent hybridization probes to evaluate codon 482, a known site of mutations in BCRP mRNA. In vitro cell viability and apoptosis were examined after 24 h exposure to flavopiridol. Results: BCRP mRNA expression varied over a 200-fold range. In the blast cell samples with BCRP mRNA expression > 10,000 copies/pg β-actin (n = 9), BCRP mRNA correlated proportionally with cell viability in the presence of 250 nM flavopiridol (r = 0.86, P = 0.003) and with apoptosis induced by flavopiridol (r = 0.71, P = 0.031). In contrast, MDR1 mRNA expression did not correlate with either flavopiridol cytotoxicity or induction of apoptosis. Melting point analysis of the hybridization probes determined that all 21 patient samples had arginine at codon 482 of BCRP mRNA, the wild-type form. Conclusions: These results suggest that unlike P-gp, BCRP may play a role in leukemia cellular resistance to flavopiridol. No mutations at codon 482 were observed in BCRP mRNA in this group of patients.

AB - Purpose: Flavopiridol is a cyclin-dependent kinase inhibitor currently undergoing human clinical trials. As clinical development is pursued, it becomes important to evaluate resistance mechanisms to flavopiridol. To elucidate the contribution of breast cancer resistance protein (BCRP) to cellular resistance to flavopiridol in acute myeloid leukemia, we studied the relationship between cellular resistance to flavopiridol and mRNA expression of BCRP or P-glycoprotein (P-gp, product of MDR1 gene) in blast cells from adult patients with acute leukemia. Experimental Design: Twenty-one blast cell samples from 20 patients were studied. The expression of BCRP, P-gp, β-actin mRNA was determined by real-time reverse transcription-PCR, using fluorescent hybridization probes to evaluate codon 482, a known site of mutations in BCRP mRNA. In vitro cell viability and apoptosis were examined after 24 h exposure to flavopiridol. Results: BCRP mRNA expression varied over a 200-fold range. In the blast cell samples with BCRP mRNA expression > 10,000 copies/pg β-actin (n = 9), BCRP mRNA correlated proportionally with cell viability in the presence of 250 nM flavopiridol (r = 0.86, P = 0.003) and with apoptosis induced by flavopiridol (r = 0.71, P = 0.031). In contrast, MDR1 mRNA expression did not correlate with either flavopiridol cytotoxicity or induction of apoptosis. Melting point analysis of the hybridization probes determined that all 21 patient samples had arginine at codon 482 of BCRP mRNA, the wild-type form. Conclusions: These results suggest that unlike P-gp, BCRP may play a role in leukemia cellular resistance to flavopiridol. No mutations at codon 482 were observed in BCRP mRNA in this group of patients.

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