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Research ArticleArticle

Identification of Cytoskeletal [14C]Carboplatin-Binding Proteins Reveals Reduced Expression and Disorganization of Actin and Filamin in Cisplatin-Resistant Cell Lines

Ding-Wu Shen, Xing-Jie Liang, Mary Ann Gawinowicz and Michael M. Gottesman
Molecular Pharmacology October 2004, 66 (4) 789-793; DOI: https://doi.org/10.1124/mol.66.4.789
Ding-Wu Shen
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Xing-Jie Liang
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Mary Ann Gawinowicz
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Michael M. Gottesman
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Abstract

Cisplatin resistant (CP-r) cells often show decreased uptake of cisplatin in association with reduced cell surface proteins and decreased endocytosis. In this report, two major [14C]carboplatin-binding proteins were identified as filamin and actin by photoaffinity labeling and mass spectrometry. Decreased expression of these two proteins was found in two different human CP-r cell lines (KB-CP20 and 7404-CP20), in comparison with their parental cell lines (KB-3-1 and BEL-7404), respectively. Disorganization of β-actin and filamin 250 and 90 was also detected in these CP-r cells by confocal microscopy. Transfection of a wild-type actin-enhanced green fluorescent protein (EGFP) expression vector into 7404-CP20 cells resulted in a nonfilamentous actin-EGFP distribution compared with a normal distribution in the cisplatin-sensitive BEL-7404 cells, suggesting that cytoskeletal organization is disturbed in the CP-r cells. The identification of actin and filamin as [14C]carboplatin-binding proteins and decreased expression and disorganization of several cytoskeletal proteins in CP-r cells provide a molecular and cellular basis for the known defect in endocytosis in these cells.

  • Received January 27, 2004.
  • Accepted June 10, 2004.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 66 (4)
Molecular Pharmacology
Vol. 66, Issue 4
1 Oct 2004
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Research ArticleArticle

Identification of Cytoskeletal [14C]Carboplatin-Binding Proteins Reveals Reduced Expression and Disorganization of Actin and Filamin in Cisplatin-Resistant Cell Lines

Ding-Wu Shen, Xing-Jie Liang, Mary Ann Gawinowicz and Michael M. Gottesman
Molecular Pharmacology October 1, 2004, 66 (4) 789-793; DOI: https://doi.org/10.1124/mol.66.4.789

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Research ArticleArticle

Identification of Cytoskeletal [14C]Carboplatin-Binding Proteins Reveals Reduced Expression and Disorganization of Actin and Filamin in Cisplatin-Resistant Cell Lines

Ding-Wu Shen, Xing-Jie Liang, Mary Ann Gawinowicz and Michael M. Gottesman
Molecular Pharmacology October 1, 2004, 66 (4) 789-793; DOI: https://doi.org/10.1124/mol.66.4.789
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