Effect of 6-O-α-maltosyl-β cyclodextrin and its cholesterol inclusion complex on cellular cholesterol levels and ABCA1 and ABCG1 expression in mouse mastocytoma P-815 cells.
We have previously described 6-O-α-maltosyl-β cyclodextrin (Mal-βCD), which forms soluble inclusion complex with cholesterol. Here we further investigated the effect of Mal-βCD and cholesterol/Mal-βCD inclusion complex (CLM) on cellular cholesterol levels in a mouse mast cell line, mastocytoma P-815 cells (P-815 cells). Mal-βCD removes cellular cholesterol forming inclusion complexes, while Mal-βCD-induced lack of cellular cholesterol was replenished by the addition of CLM without cytotoxicity. Reduction and replenishment of cellular cholesterol in Mal-βCD- and/or CLM-treated P-815 cells, respectively, were demonstrated by LC/MS and fluorescence microscopy with filipin III. CLM rather than free Mal-βCD and free cholesterol was efficiently incorporated into P-815 cells and its incorporation was inhibited by incubation at low temperature, or with sodium azide and cytochalasin D. P-815 cells have been confirmed to express ATP-binding cassette (ABC) transporters, ABCA1, ABCG1, and P-glycoprotein (P-gp), by Western blot and mRNA analysis. Cholesterol reduction by Mal-βCD abolishes the mRNA and protein expression of ABCA1 and ABCG1, but not of P-gp. Cholesterol loading by CLM restores the diminished ABCA1 and ABCG1 mRNA expression in Mal-βCD-treated P-815 cells. However, both Mal-βCD and CLM had no effect on P-gp activity measured by the rhodamine 123 efflux assay. These results indicate that alteration of cholesterol levels with Mal-βCD or CLM led to down- or up-regulation of ABCA1 and ABCG1 expression in P-815 cells.
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, Institute for Biosciences, Mukogawa Women's University, 11-68 Koshien-Kyubancho, Nishinomiya-shi, Hyogo 663-8179, Japan. okada@mukogawa-u.ac.jp
Yasuyo
Y
Okada
Kiyomi
K
Ueyama
Jyun-ichi
J
Nishikawa
Masanori
M
Semma
Atsushi
A
Ichikawa
D064286:ATP Binding Cassette Transporter 1; D000070998:ATP Binding Cassette Transporter, Subfamily G, Member 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000378:metabolism* / Q000494:pharmacology* / Q000633:toxicity; D004705:Endocytosis / Q000187:drug effects; D005372:Filipin / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D034801:Mastocytoma; D051379:Mice; D010539:Permeability; D047392:beta-Cyclodextrins / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity
C487540:ABCG1 protein, mouse; D064286:ATP Binding Cassette Transporter 1; D000070998:ATP Binding Cassette Transporter, Subfamily G, Member 1; D018528:ATP-Binding Cassette Transporters; D005456:Fluorescent Dyes; D008074:Lipoproteins; D047392:beta-Cyclodextrins; C075170:6-O-maltosyl-beta-cyclodextrin; D005372:Filipin; D002784:Cholesterol; D007770:L-Lactate Dehydrogenase