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Vol. 30, Issue 1, 4-6, January 2002

SHORT 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

Department of Hospital Pharmacy
(Ts.N, T.Sa., N.O., K.O.),
Second Department of Internal Medicine
(T.T., N.A., M.K),
First Department of Surgery
(T.K., Ta.N., Y.K.),
Department of Urology (T.Sh.),
Departments of Clinical Genetics
and International Center for
Medical Research (T.Sh., M.M.),
School of Medicine, Kobe University,
Kobe, Japan
Department of Gastroenterology
(K.I.K., S.R.K.),
Kobe Asahi Hospital, Kobe, Japan

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.

Copyright © 2002 by The American Society for Pharmacology and Experimental Therapeutics


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Copyright © 2002 by the American Society for Pharmacology and Experimental Therapeutics.