Uptake of 4-chloro-2-methylphenoxyacetic acid (MCPA) from the apical membrane of Caco-2 cells by the monocarboxylic acid transporter

Uptake of Aristolochic Acid I into Caco-2 Cells by Monocarboxylic Acid Transporters

“…As described previously, the cells were cultured on 35-mm six-well culture dishes coated with rat tail collagen type I (Becton Dickinson, Bedford, MA, U.S.A.), 9,10) or permeable membranes (Cell Culture Insert, 0.9 cm 2 growth area: Becton Dickinson) 16) in DMEM containing FBS (10%), NEAA (1%), streptomycin (100 U/mL) and penicillin G (70 µg/mL) at 37°C in a humidified atmosphere of 5% CO 2 -95% air. The culture medium was replaced three times a week, and confluent Caco-2 cell monolayers cultured for 7-9 d on the dishes or for 21 or 22 d on the membrane were used in this study.…”

“…[9][10][11] The incubation medium used for the uptake study was Hanks' balanced salt solution (HBSS) containing 25 mM D-glucose and 10 mM 2-(N-morpholino)ethanesulfonate (MES) (pH 5.5, 6.0 or 6.5) or 10 mM N-hydroxyethylpiperazine-N′-2-ethanesulfonate (HEPES) (pH 7.0, 7.4 or 8.0). The culture medium was removed, and the cells were preincubated at 37°C or 4°C for 20 min in 1.5 mL of the incubation medium at pH 7.4.…”

“…Na + -free HBSS was prepared by replacing NaCl with choline chloride, and omitting NaH 2 PO 4 , to examine whether the uptake of AAI was Na + dependent. 11) In the metabolic inhibition experiments, the cells were treated with 10 mM NaN 3 or 25 µM FCCP in the incubation medium at 37°C for 20 min, [9][10][11] prior to incubation with AAI. To investigate the uptake of AAI via specific mechanisms, the cells were coincubated with 50 µM AAI and various compounds.…”

“…The cells were suspended in 1.0 mL of extraction solution (1 N H 3 PO 4 : methanol= 1 : 1) for 60 min at room temperature, and the cells were scraped off and collected using a cell scraper. [9][10][11] The suspension was centrifuged at 13000×g for 10 min, and a 50-µL aliquot of the supernatant was injected onto the HPLC system. Transport Experiments The transport experiment was performed as described previously.…”

“…5) MCTs have an important role in the intestinal absorption of not only short-chain monocarboxylic acids, such as L-lactic acid, propionic acid and pyruvic acid, but also exogenous monocarboxylic acid compounds, such as benzoic acid, ferulic acid, nonsteroidal anti-inflammatory drugs and phenoxyacetic acid herbicides. [6][7][8][9][10][11] The human colorectal adenocarcinomal cell line Caco-2 is a useful model with which to study the intestinal absorption of various compounds as the cells are morphologically and functionally similar to human small intestinal epithelial cells. 12) Caco-2 cells express various MCT subtypes such as MCT1, MCT3, MCT4, MCT5 and MCT6.…”