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Rapid CommunicationShort Communication

Real-Time Quantitative Polymerase Chain Reaction for MDR1, MRP1, MRP2, and CYP3A-mRNA Levels in Caco-2 Cell Lines, Human Duodenal Enterocytes, Normal Colorectal Tissues, and Colorectal Adenocarcinomas

Tsutomu Nakamura, Toshiyuki Sakaeda, Nobuko Ohmoto, Takao Tamura, Nobuo Aoyama, Toshiro Shirakawa, Takashi Kamigaki, Takeshi Nakamura, Ke Ih Kim, Soo Ryang Kim, Yoshikazu Kuroda, Masafumi Matsuo, Masato Kasuga and Katsuhiko Okumura
Drug Metabolism and Disposition January 2002, 30 (1) 4-6; DOI: https://doi.org/10.1124/dmd.30.1.4
Tsutomu Nakamura
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Toshiyuki Sakaeda
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Nobuko Ohmoto
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Takao Tamura
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Nobuo Aoyama
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Toshiro Shirakawa
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Takashi Kamigaki
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Takeshi Nakamura
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Ke Ih Kim
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Soo Ryang Kim
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Yoshikazu Kuroda
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Masafumi Matsuo
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Masato Kasuga
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Katsuhiko Okumura
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Abstract

The expression levels of mRNAs for MDR1 (P-glycoprotein), multidrug resistance-associated proteins (MRP1, MRP2), and cytochrome P450 3A (CYP3A) in Caco-2 cells were quantitatively compared with those in human duodenal enterocytes, normal colorectal tissues, and colorectal adenocarcinomas. Caco-2 cells (passages 36–88) were kindly supplied by several laboratories in Japan. Human duodenal enterocytes were obtained from five healthy male volunteers. Normal colorectal tissues and colorectal adenocarcinomas were simultaneously obtained from seven patients with primary colorectal adenocarcinoma. MDR1, MRP1, MRP2, and CYP3A mRNA levels were determined by real-time quantitative polymerase chain reactions (PCR). Relative concentrations of mRNAs for target proteins (MDR1, MRP1, MRP2, and CYP3A) and glyceraldehyde-3-phosphate dehydrogenase in Caco-2 cells were 1.00 ± 0.15, 1.02 ± 0.06, 0.94 ± 0.10, and 0.68 ±0.60, respectively, and those in human enterocytes were about 12-, 3-, 7-, and 8000-fold higher than in the Caco-2 cells, respectively. In contrast, MDR1, MRP1, and CYP3A mRNA levels in Caco-2 cells were comparable to those in normal colorectal tissue and colorectal adenocarcinoma.

Footnotes

  • Abbreviations used are::
    MRP
    multidrug resistance-associated protein
    RT-PCR
    reverse transcription-polymerase chain reaction
    GAPDH
    glyceraldehyde-3-phosphate dehydrogenase
    CT
    threshold cycle number
    • Received June 25, 2001.
    • Accepted October 16, 2001.
  • The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition: 30 (1)
Drug Metabolism and Disposition
Vol. 30, Issue 1
1 Jan 2002
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Rapid CommunicationShort Communication

Real-Time Quantitative Polymerase Chain Reaction for MDR1, MRP1, MRP2, and CYP3A-mRNA Levels in Caco-2 Cell Lines, Human Duodenal Enterocytes, Normal Colorectal Tissues, and Colorectal Adenocarcinomas

Tsutomu Nakamura, Toshiyuki Sakaeda, Nobuko Ohmoto, Takao Tamura, Nobuo Aoyama, Toshiro Shirakawa, Takashi Kamigaki, Takeshi Nakamura, Ke Ih Kim, Soo Ryang Kim, Yoshikazu Kuroda, Masafumi Matsuo, Masato Kasuga and Katsuhiko Okumura
Drug Metabolism and Disposition January 1, 2002, 30 (1) 4-6; DOI: https://doi.org/10.1124/dmd.30.1.4

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Rapid CommunicationShort Communication

Real-Time Quantitative Polymerase Chain Reaction for MDR1, MRP1, MRP2, and CYP3A-mRNA Levels in Caco-2 Cell Lines, Human Duodenal Enterocytes, Normal Colorectal Tissues, and Colorectal Adenocarcinomas

Tsutomu Nakamura, Toshiyuki Sakaeda, Nobuko Ohmoto, Takao Tamura, Nobuo Aoyama, Toshiro Shirakawa, Takashi Kamigaki, Takeshi Nakamura, Ke Ih Kim, Soo Ryang Kim, Yoshikazu Kuroda, Masafumi Matsuo, Masato Kasuga and Katsuhiko Okumura
Drug Metabolism and Disposition January 1, 2002, 30 (1) 4-6; DOI: https://doi.org/10.1124/dmd.30.1.4
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