Gene expression profiles of ABC transporters and cytochrome P450 3A in Caco-2 and human colorectal cancer cell lines.
OBJECTIVE
The mRNA levels of MDR1 (P-glycoprotein), multidrug resistance-associated proteins (MRP1, MRP2), cytochrome P450 3A (CYP3A) and villin in human colorectal cell lines (HCT-15, LoVo DLD-1, HCT-116 and SW620) were quantitatively compared with those in Caco-2 cells.


METHODS
The mRNA levels were determined by real time quantitative polymerase chain reaction and expressed as the relative concen trations of MDR1 mRNA to glyceraldehyde-3-phosphate dehydro genase (GAPDH) mRNA. RESULTSl MDR1 mRNA was expressed in HCT-15 LoVo and DLD-1 cells at similar or lower level to Caco-2. The expression of MRP mRNA in the cell lines tested was comparable with Caco-2. MRP. mRNA was detected only in HCT-116 and SW620 at significantly lower level than Caco-2. CYP3A mRNA was detected in HCT-15 LoVo, DLD-1 and SW620 at similar level to Caco-2.


CONCLUSIONS
HCT-15 LoVo and DLD-1 cells express proteins important for regulating the intestinal absorption of drugs, i.e., MDR1 MRP1 and CYP3A, whereas HCT-116 and SW620 cells were not acceptable for evaluation of absorption properties of drug candidates
Nakumura
Tsutomu
T
Department of Hospital Pharmacy, School of Medicine, Kobe University, Chuo-ku, Kobe 650-0017, Japan.
Sakaeda
Toshiyuki
T
Ohmoto
Nobuko
N
Moriya
Yuka
Y
Komoto
Chiho
C
Shirakawa
Toshiro
T
Gotoh
Akinobu
A
Matsuo
Masafumi
M
Okmura
Katsuhiko
K
D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis* / Q000235:genetics; D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis* / Q000235:genetics; D018938:Caco-2 Cells; D015179:Colorectal Neoplasms / Q000201:enzymology / Q000378:metabolism*; D051544:Cytochrome P-450 CYP3A; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured
D018528:ATP-Binding Cassette Transporters; D012333:RNA, Messenger; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; D010089:Oxidoreductases, N-Demethylating