title: Contribution of aquaporin 9 and multidrug resistance-associated protein 2 to differential sensitivity to arsenite between primary cultured chorion and amnion cells prepared from human fetal membranes.
abstract: Arsenic trioxide (arsenite, As(III)) has shown a remarkable clinical efficacy, whereas its side effects are still a serious concern. Therefore, it is critical to understand the effects of As(III) on human-derived normal cells for revealing the mechanisms underlying these side effects. We examined the effects of As(III) on primary cultured chorion (C) and amnion (A) cells prepared from human fetal membranes. A significant dose-dependent As(III)-mediated cytotoxicity was observed in the C-cells accompanied with an increase of lactate dehydrogenase (LDH) release. Higher concentrations of As(III) were required for the A-cells to show cytotoxicity and LDH release, suggesting that the C-cells were more sensitive to As(III) than the A-cells. The expression levels of aquaporin 9 (AQP9) were approximately 2 times higher in the C-cells than those in the A-cells. Both intracellular arsenic accumulation and its cytotoxicity in the C-cells were significantly abrogated by sorbitol, a competitive AQP9 inhibitor, in a dose-dependent manner. The protein expression levels of multidrug resistance-associated protein (MRP) 2 were downregulated by As(III) in the C-cells, but not in the A-cells. No significant differences in the expression levels of MRP1 were observed between C- and A-cells. The protein expression of P-glycoprotein (P-gp) was hardly detected in both cells, although a detectable amount of its mRNA was observed. Cyclosporine A, a broad-spectrum inhibitor for ABC transporters, and MK571, a MRP inhibitor, but not PGP-4008, a P-gp specific inhibitor, potently sensitized both cells to As(III)-mediated cytotoxicity. These results suggest that AQP9 and MRP2 are involved in controlling arsenic accumulation in these normal cells, which then contribute to differential sensitivity to As(III) cytotoxicity between these cells.
authors.affiliation: Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy & Life Sciences, Horinouchi, Hachioji, Tokyo, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Yoshino
authors.forename: Bo
authors.initials: B
authors.lastname: Yuan
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Kaise
authors.forename: Makoto
authors.initials: M
authors.lastname: Takeichi
authors.forename: Sachiko
authors.initials: S
authors.lastname: Tanaka
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Hirano
authors.forename: Deanna L
authors.initials: DL
authors.lastname: Kroetz
authors.forename: Hiroo
authors.initials: H
authors.lastname: Toyoda
mesh_terms: D000650:Amnion / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020346:Aquaporins / Q000502:physiology*; D018053:Arsenites / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002823:Chorion / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005321:Extraembryonic Membranes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis*
chemical_list: D020346:Aquaporins; D018053:Arsenites; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C000715087:ABCC2 protein, human; C109669:AQP9 protein, human; C015001:arsenite