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Polymorphisms and Methylation of the Reduced Folate Carrier in Osteosarcoma

  • Symposium: Molecular Genetics in Sarcoma
  • Published:
Clinical Orthopaedics and Related Research

Abstract

High-dose methotrexate is a standard component in the treatment of osteogenic sarcoma. Impaired methotrexate uptake associated with decreased reduced folate carrier expression is a common mechanism of methotrexate resistance in osteogenic sarcoma samples. We investigated whether promoter methylation and polymorphisms in the 3′ untranslated region are involved in regulating reduced folate carrier expression. In a cohort of 66 osteogenic sarcoma specimens, quantitative methylation-specific polymerase chain reaction and single-strand conformation polymorphism were performed. We found detectable levels of promoter methylation in 84.3% of samples. When related to the reduced folate carrier mRNA levels, a trend was observed that reduced folate carrier expression is lower in samples (median, 0.7) with greater than 10% DNA methylation as compared with those (median, 2.3) with less than 10% DNA methylation. The heterozygous polymorphisms of 2582 T/G and 2617C/T in the 3′ untranslated region showed reduced folate carrier expression (median, 0.9) as compared with the wild-type 2582T and 2617C (median, 4.2). The data suggest promoter methylation and polymorphisms in the 3′ untranslated region of the reduced folate carrier may be involved in its transcriptional regulation in osteogenic sarcoma. Further study is required to confirm this finding.

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Acknowledgments

We thank Rebecca Sowers, BS, and BethAnne Mazza, BS, for the technical support provided in this laboratory. We also thank Dr Joseph R. Bertino and Dr Debabrata Banerjee of the Cancer Institute of New Jersey, New Brunswick, NJ, for the fruitful discussions and insights. We are in debt to Dr Andrew G. Huvos of MSKCC, who is forever in our memory. Statistical analysis was performed by the Department of Epidemiology and Biostatistics (JQ) at MSKCC.

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Author notes

  1. Jing Qin PhD

    Present address: Department of Health & Human Services, Public Health Service, National Institute of Health, Bethesda, MD, USA

  2. Bang H. Hoang MD

    Present address: Department of Orthopaedic Surgery, University of California, Irvine, CA, USA

Authors and Affiliations

  1. Department of Pediatrics and Molecular Pharmacology, The Albert Einstein College of Medicine, The Children’s Hospital at Montefiore, 3415 Bainbridge Avenue, Rosenthal 3rd Floor, Bronx, NY, 10467, USA

    Rui Yang MD & Richard Gorlick MD

  2. Department of Epidemiology and Biostatistics, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

    Jing Qin PhD

  3. Orthopedic Surgery Service, Weill Medical College of Cornell, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

    Bang H. Hoang MD & John H. Healey MD

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  1. Rui Yang MD
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  2. Jing Qin PhD
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  3. Bang H. Hoang MD
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  4. John H. Healey MD
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  5. Richard Gorlick MD
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Correspondence to Richard Gorlick MD.

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Each author certifies that he or she has no commercial associations (eg, consultancies, stock ownership, equity interest, patent/licensing arrangements, etc) that might pose a conflict of interest in connection with the submitted article.

Each author certifies that his or her institution has approved the human protocol for this investigation, that all investigations were conducted in conformity with ethical principles of research, and that informed consent for participation in the study was obtained.

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Yang, R., Qin, J., Hoang, B.H. et al. Polymorphisms and Methylation of the Reduced Folate Carrier in Osteosarcoma. Clin Orthop Relat Res 466, 2046–2051 (2008). https://doi.org/10.1007/s11999-008-0323-3

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  • DOI: https://doi.org/10.1007/s11999-008-0323-3

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