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Anticancer Res ; 30(9): 3661-8, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20944151

RESUMO

Overexpression of P-glycoprotein (P-gp), a plasma membrane drug transporter (ABCB1, a member of the ABC transporter family), is the most prevalent cause of multidrug resistance in cancer tissues. Lectin concanavalin A (ConA) induces massive cell death of L1210 leukemia cells (S). Cell sublines of L1210 in which P-gp overexpression was induced by selection with vincristine (R) or by stable transfection with a plasmid encoding full-length human P-gp (T) were less sensitive to ConA. Both P-gp-positive cell lines exhibited typical P-gp-mediated multidrug resistance. Resistance of R and T cells to ConA was associated with lower binding of ConA as compared to S cells when analysed by the following methods: (i) SDS PAGE and electroblotting of proteins in the crude membrane fraction followed by detection with biotinylated ConA and avidin-peroxidase, and (ii) fluorescent cytometry or confocal microscopy of the intact cells with surfaces labeled by FITC-ConA. These data indicated that the presence of P-glycoprotein in L1210 cells independently of the mode of its expression induced down-regulation of cell surface saccharide targets of ConA. Therefore, this feature may be considered as a secondary cellular response to P-glycoprotein expression.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Membrana Celular/metabolismo , Concanavalina A/metabolismo , Resistencia a Medicamentos Antineoplásicos , Animais , Linhagem Celular Tumoral , Separação Celular , Regulação para Baixo , Eletroforese em Gel de Poliacrilamida , Citometria de Fluxo , Glicoproteínas/metabolismo , Humanos , Immunoblotting , Camundongos , Microscopia Confocal , Transfecção
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