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Research ArticleCELLULAR AND MOLECULAR

Transporter Gene Expression in Lactating and Nonlactating Human Mammary Epithelial Cells Using Real-Time Reverse Transcription-Polymerase Chain Reaction

J. Alcorn, X. Lu, J. A. Moscow and P. J. McNamara
Journal of Pharmacology and Experimental Therapeutics November 2002, 303 (2) 487-496; DOI: https://doi.org/10.1124/jpet.102.038315
J. Alcorn
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X. Lu
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J. A. Moscow
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P. J. McNamara
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Abstract

Transporter-mediated processes in the lactating mammary gland may explain the significant accumulation of certain drugs in breast milk. The purpose of this study was to identify potential candidate drug transport proteins involved in drug accumulation in milk. Quantitative reverse transcription-polymerase chain reaction methods were developed to determine the relative RNA levels of 30 different drug transporter genes. Transporter gene RNA levels in lactating mammary epithelial cells (MEC) purified from pooled fresh breast milk samples were compared with levels in nonlactating MEC, liver, and kidney tissue. Transcripts were detected in lactating MEC for OCT1, OCT3, OCTN1, OCTN2, OATP-A, OATP-B, OATP-D, OATP-E, MRP1, MRP2, MRP5, MDR1, CNT1, CNT3, ENT1, ENT3, NCBT1, PEPT1, and PEPT2. No transcripts were detected for OCT2, OAT1, OAT2, OAT3, OAT4, OATP-C, MRP3, MRP4, CNT2, ENT2, and NCBT2. Lactating MEC demonstrated more than 4-fold higher RNA levels of OCT1, OCTN1, PEPT2, CNT1, CNT3, and ENT3, and more than 4-fold lower RNA levels of MDR1 and OCTN2 relative to nonlactating MEC. Lactating MEC showed significantly higher RNA levels of CNT3 relative to liver and kidney, increased PEPT2 RNA levels relative to liver, and increased OATP-A RNA levels relative to kidney. These data imply CNT3 may play a specialized role in nucleoside accumulation in milk and may identify an important role for PEPT2 and OATP-A transporters at the lactating mammary epithelium. Furthermore, transporters expressed in lactating MEC identify a potential role for these transporters in drug disposition at the mammary gland.

Footnotes

  • The University of Kentucky Research Challenge Trust Fund provided financial support for J.A. This work was supported by National Institutes of Health Grant HD37463.

  • DOI: 10.1124/jpet.102.038315

  • Abbreviations:
    M/S
    milk drug concentration to serum drug concentration ratio
    OCT
    organic cation transporter
    OAT
    organic anion transporter
    OATP
    organic anion transporting polypeptide
    MRP
    multidrug resistance-associated protein
    MDR
    multidrug resistance transporter
    MEC
    mammary epithelial cells
    RT-PCR
    reverse transcription-polymerase chain reaction
    FBS
    fetal bovine serum
    PBS
    phosphate-buffered saline
    EMA
    epithelial membrane antigen
    MPC
    magnetic particle concentrator
    CALLA
    common acute lymphoblastic leukemia antigen
    CNT
    concentrative nucleoside transporter
    ENT
    equilibrative nucleoside transporter
    NCBT
    nucleobase transporter
    OCTN
    organic cation/carnitine transporter
    PEPT
    oligopeptide transporter
    • Received May 2, 2002.
    • Accepted June 25, 2002.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 303 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 303, Issue 2
1 Nov 2002
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Research ArticleCELLULAR AND MOLECULAR

Transporter Gene Expression in Lactating and Nonlactating Human Mammary Epithelial Cells Using Real-Time Reverse Transcription-Polymerase Chain Reaction

J. Alcorn, X. Lu, J. A. Moscow and P. J. McNamara
Journal of Pharmacology and Experimental Therapeutics November 1, 2002, 303 (2) 487-496; DOI: https://doi.org/10.1124/jpet.102.038315

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Research ArticleCELLULAR AND MOLECULAR

Transporter Gene Expression in Lactating and Nonlactating Human Mammary Epithelial Cells Using Real-Time Reverse Transcription-Polymerase Chain Reaction

J. Alcorn, X. Lu, J. A. Moscow and P. J. McNamara
Journal of Pharmacology and Experimental Therapeutics November 1, 2002, 303 (2) 487-496; DOI: https://doi.org/10.1124/jpet.102.038315
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