Changes in adsorption and permeability of mitoxantrone on plasma membrane of BCRP/MXR resistant cells.
A selective analysis of adsorbed mitoxantrone (MTX) was performed by surface-enhanced Raman scattering (SERS) at the range of cellular membrane. Disruption of the membrane fluidity was carried out to appraise changes in membrane adsorption of MTX and drug uptake in sensitive (HCT-116 S) and resistant BCRP/MXR (HCT-116 R) cells. Based on spectral MTX modifications, micro-SERS spectroscopy discriminated clearly drug adsorption phenomena on plasma membrane from drug in solution. A 3-fold higher SERS intensity of MTX for HCT-116 R was observed concluding to a higher drug adsorption on resistant membrane. The increase of membrane fluidity with benzyl alcohol (BA) or chloroform (CF) resulted in a 3-fold decrease of MTX adsorption on HCT-116 R, exclusively. BA and CF improved intracellular accumulation of MTX (e.g., 823 and 191 pmol MTX/10(6) HCT-116 R incubated with or without BA). At 4 degrees C, drug accumulation measurements showed a decrease of MTX permeability in resistant membrane (42 pmol MTX/10(6) cells), restored with fluidizers (e.g., 342 pmol MTX/10(6) cells with BA). Fluorescence confocal microscopy involved an exclusive MTX emission around the plasma membrane of resistant cells whereas fluidizers increased the intracellular uptake of MTX in both cell lines at the same time with less drug emission around the plasma membrane. Changes of the membrane structure of resistant cells should modify both drug adsorption and membrane permeation.
Breuzard
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G
Unité MéDIAN CNRS UMR 6142, IFR 53, UFR de pharmacie, 51 rue Cognacq-Jay, 51096 Reims Cedex, France.
Piot
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Angiboust
J-F
JF
Manfait
M
M
Candeil
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L
Del Rio
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Millot
J-M
JM
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000327:Adsorption / Q000187:drug effects; D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism*; D002725:Chloroform / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance; D019008:Drug Resistance, Neoplasm; D006801:Humans; D018613:Microscopy, Confocal; D008942:Mitoxantrone / Q000494:pharmacology*; D009363:Neoplasm Proteins / Q000378:metabolism; D010539:Permeability / Q000187:drug effects; D013053:Spectrophotometry; D013696:Temperature; D013997:Time Factors
C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; D009363:Neoplasm Proteins; D002725:Chloroform; D008942:Mitoxantrone