Cellular folate status modulates the expression of BCRP and MRP multidrug transporters in cancer cell lines from different origins

As cellular folate levels seem to have a different effect on cancer cells from different origins, we extended our initial study to a broader panel of cancer cells. BCRP and MRP1-5 expression was determined in KB, OVCAR-3, IGROV-1, ZR75-1/R/MTX, SCC-11B, SCC-22B, and WiDr either grown in standard RPMI 1640 containing 2.3 μmol/L supraphysiologic concentration of folic acid [high folate (HF)] or adapted to more physiologic concentrations [1-5 nmol/L folic acid or leucovorin; low folate (LF)]. Compared with the HF counterparts, KB LF cells displayed 16.1-fold increased MRP3 and OVCAR-3 LF cells showed 4.8-fold increased MRP4 mRNA levels along with increased MRP3 and MRP4 protein expression, respectively. A marked increase on BCRP protein and mRNA expression was observed in WiDr LF cells. These cells acquired ∼2-fold resistance to mitoxantrone compared with the HF cell line, a phenotype that could be reverted by the BCRP inhibitor Ko143. Of note, WiDr cells expressed BCRP in the intracellular compartment, similarly to what we have described for Caco-2 cells. Our results provide further evidence for an important role of cellular folate status in the modulation of the expression of multidrug resistance transporters in cancer cells. We show that up-regulation of intracellularly localized BCRP in response to adaptation to LF conditions may be a common feature within a panel of colon cancer cell lines. Under these circumstances, folate supplementation might improve the efficacy of chemotherapeutic drugs by decreasing BCRP expression. [Mol Cancer Ther 2009;8(3):655–64]

[1]  E. Giovannetti,et al.  Folate deprivation induces BCRP (ABCG2) expression and mitoxantrone resistance in Caco‐2 cells , 2008, International journal of cancer.

[2]  G. Jansen,et al.  Drug transporters: recent advances concerning BCRP and tyrosine kinase inhibitors , 2008 .

[3]  M. Ratnam,et al.  The folate receptor: What does it promise in tissue-targeted therapeutics? , 2007, Cancer and Metastasis Reviews.

[4]  M. Akabas,et al.  Identification of an Intestinal Folate Transporter and the Molecular Basis for Hereditary Folate Malabsorption , 2006, Cell.

[5]  M. Schell,et al.  ABCG2 expression, function, and promoter methylation in human multiple myeloma. , 2006, Blood.

[6]  G. Szakács,et al.  Human multidrug resistance ABCB and ABCG transporters: participation in a chemoimmunity defense system. , 2006, Physiological reviews.

[7]  S. Bates,et al.  Aberrant Promoter Methylation of the ABCG2 Gene in Renal Carcinoma , 2006, Molecular and Cellular Biology.

[8]  Y. Assaraf.  The role of multidrug resistance efflux transporters in antifolate resistance and folate homeostasis. , 2006, Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy.

[9]  S. Cole,et al.  Transmembrane transport of endo- and xenobiotics by mammalian ATP-binding cassette multidrug resistance proteins. , 2006, Physiological reviews.

[10]  R. Pieters,et al.  Multidrug resistance proteins and folate supplementation: therapeutic implications for antifolates and other classes of drugs in cancer treatment , 2006, Cancer Chemotherapy and Pharmacology.

[11]  M. Gottesman,et al.  The molecular basis of multidrug resistance in cancer: The early years of P‐glycoprotein research , 2006, FEBS letters.

[12]  Y. Assaraf,et al.  Novel extracellular vesicles mediate an ABCG2-dependent anticancer drug sequestration and resistance. , 2005, Cancer research.

[13]  S. Caudill,et al.  Biochemical indicators of B vitamin status in the US population after folic acid fortification: results from the National Health and Nutrition Examination Survey 1999-2000. , 2005, The American journal of clinical nutrition.

[14]  Elisa Giovannetti,et al.  Cellular and Pharmacogenetics Foundation of Synergistic Interaction of Pemetrexed and Gemcitabine in Human Non–Small-Cell Lung Cancer Cells , 2005, Molecular Pharmacology.

[15]  G. Peters,et al.  Quantitative real time PCR of deoxycytidine kinase mRNA by Light Cycler PCR in relation to enzyme activity and gemcitabine sensitivity. , 2004, Cancer letters.

[16]  P. Wielinga,et al.  Mrp4 Confers Resistance to Topotecan and Protects the Brain from Chemotherapy , 2004, Molecular and Cellular Biology.

[17]  B. Dijkmans,et al.  Sulfasalazine is a potent inhibitor of the reduced folate carrier: implications for combination therapies with methotrexate in rheumatoid arthritis. , 2004, Arthritis and rheumatism.

[18]  Yehuda G. Assaraf,et al.  Folate Deprivation Results in the Loss of Breast Cancer Resistance Protein (BCRP/ABCG2) Expression , 2004, Journal of Biological Chemistry.

[19]  P. Stover.  Physiology of folate and vitamin B12 in health and disease. , 2004, Nutrition reviews.

[20]  M. Ratnam,et al.  Distribution, functionality and gene regulation of folate receptor isoforms: implications in targeted therapy. , 2004, Advanced drug delivery reviews.

[21]  G. Peters,et al.  Folate concentration dependent transport activity of the Multidrug Resistance Protein 1 (ABCC1). , 2004, Biochemical pharmacology.

[22]  B. Dijkmans,et al.  Development of sulfasalazine resistance in human T cells induces expression of the multidrug resistance transporter ABCG2 (BCRP) and augmented production of TNFα , 2004, Annals of the rheumatic diseases.

[23]  L. Doyle,et al.  Multidrug resistance mediated by the breast cancer resistance protein BCRP (ABCG2) , 2003, Oncogene.

[24]  S. Simon,et al.  Subcellular localization and activity of multidrug resistance proteins. , 2003, Molecular biology of the cell.

[25]  H. Said,et al.  Folate uptake in the human intestine: Promoter activity and effect of folate deficiency , 2003, Journal of cellular physiology.

[26]  Claude Denham,et al.  Phase III study of pemetrexed in combination with cisplatin versus cisplatin alone in patients with malignant pleural mesothelioma. , 2003, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[27]  G. Peters,et al.  Expression of deoxycytidine kinase in leukaemic cells compared with solid tumour cell lines, liver metastases and normal liver. , 2003, European journal of cancer.

[28]  G. Peters,et al.  The role of multidrug resistance proteins MRP1, MRP2 and MRP3 in cellular folate homeostasis. , 2003, Biochemical pharmacology.

[29]  G. Peters,et al.  Loss of Multidrug Resistance Protein 1 Expression and Folate Efflux Activity Results in a Highly Concentrative Folate Transport in Human Leukemia Cells* , 2003, The Journal of Biological Chemistry.

[30]  H. Rosing,et al.  The breast cancer resistance protein protects against a major chlorophyll-derived dietary phototoxin and protoporphyria , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[31]  W. Verschuren,et al.  Dietary monoglutamate and polyglutamate folate are associated with plasma folate concentrations in Dutch men and women aged 20-65 years. , 2002, The Journal of nutrition.

[32]  J. Schellens,et al.  Potent and specific inhibition of the breast cancer resistance protein multidrug transporter in vitro and in mouse intestine by a novel analogue of fumitremorgin C. , 2002, Molecular cancer therapeutics.

[33]  G. Jansen,et al.  Folate homeostasis and antiproliferative activity of folates and antifolates. , 2001, Nutrition.

[34]  M. Vijver,et al.  Subcellular localization and distribution of the breast cancer resistance protein transporter in normal human tissues. , 2001, Cancer research.

[35]  G. Rampersaud,et al.  Genomic DNA methylation decreases in response to moderate folate depletion in elderly women. , 2000, The American journal of clinical nutrition.

[36]  G. Peters,et al.  Folate depletion increases sensitivity of solid tumor cell lines to 5‐fluorouracil and antifolates , 2000, International journal of cancer.

[37]  M. Kool,et al.  Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies. , 2000, Cancer research.

[38]  I. Pogribny,et al.  Moderate folate depletion increases plasma homocysteine and decreases lymphocyte DNA methylation in postmenopausal women. , 1998, The Journal of nutrition.

[39]  S. B. Gates,et al.  Dietary folate and folylpolyglutamate synthetase activity in normal and neoplastic murine tissues and human tumor xenografts. , 1996, Biochemical pharmacology.

[40]  M. Flens,et al.  Tissue distribution of the multidrug resistance protein. , 1996, The American journal of pathology.

[41]  G. Peters,et al.  Screening of colon tumor cells and tissues for folylpolyglutamate synthetase activity. , 1995, Oncology research.

[42]  C. Meijer,et al.  Immunochemical detection of the multidrug resistance-associated protein MRP in human multidrug-resistant tumor cells by monoclonal antibodies. , 1994, Cancer research.

[43]  J. Schornagel,et al.  In vitro activity of novel antifolates against human squamous carcinoma cell lines of the head and neck with inherent resistance to methotrexate , 1992, International journal of cancer.

[44]  J. C. Schmitz,et al.  Pharmacokinetics of leucovorin metabolites in human plasma as a function of dose administered orally and intravenously. , 1991, Journal of the National Cancer Institute.

[45]  G. Peters,et al.  Comparison of the sulforhodamine B protein and tetrazolium (MTT) assays for in vitro chemosensitivity testing. , 1991, European journal of cancer.

[46]  J. Schornagel,et al.  Methotrexate transport in variant human CCRF-CEM leukemia cells with elevated levels of the reduced folate carrier. Selective effect on carrier-mediated transport of physiological concentrations of reduced folates. , 1990, The Journal of biological chemistry.

[47]  R. Schilsky,et al.  High-dose methotrexate: a critical reappraisal. , 1987, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[48]  T. Dale,et al.  The breast cancer resistance protein BCRP (ABCG2) concentrates drugs and carcinogenic xenotoxins into milk , 2005, Nature Medicine.

[49]  D. Goldman,et al.  Membrane transport of folates. , 2003, Vitamins and hormones.

[50]  M. Gottesman,et al.  Multidrug resistance in cancer: role of ATP–dependent transporters , 2002, Nature Reviews Cancer.