Proteomic analysis of a lymphoma-derived cell line (DG75) following treatment with a demethylating drug: modification of membrane-associated proteins.
5'-azacytidine (AZC) is a potent DNA demethylating agent used clinically for treatment of patients with malignant hemopathies. We have previously shown that AZC induces a halt in cell growth and a decrease of cell activity, without affecting cell viability. We have also shown using proteomics, that 35 polypeptides were differentially expressed in a cytoplasmic fraction. The aim of this study was to provide a more complete picture of modifications in AZC-treated cells using cell membrane preparations. Therefore the protein pattern changes following AZC treatment of the cell line DG75 were studied on a detergent-solubilized fraction obtained from these membranes. Results showed that 49 proteins were differentially expressed in the membrane fraction. Seven polypeptides were down-regulated, while 42 were up-regulated. The identity of most of these differentially expressed proteins was determined by mass spectrometry (liquid chromatography-tandem mass spectrometry or matrix-assisted laser desorption/ionization-time of flight), and the identified proteins were grouped based on cellular function and participation in biochemical and signaling pathways.
Poirier
Florence
F
Laboratoire de Biochimie des Protéines et Protéomique Immunopathologie et ImmunoIntervention, UFR SMBH Léonard de Vinci, Université Paris 13, Bobigny, France.
Joubert-Caron
Raymonde
R
Labas
Valérie
V
Caron
Michel
M
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology / Q000627:therapeutic use; D001374:Azacitidine / Q000494:pharmacology / Q000627:therapeutic use; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D005782:Gels; D006801:Humans; D008223:Lymphoma / Q000188:drug therapy / Q000378:metabolism*; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis* / Q000145:classification / Q000187:drug effects / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics; D018892:Proton-Motive Force; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000964:Antimetabolites, Antineoplastic; D005782:Gels; D008565:Membrane Proteins; D020543:Proteome; D001374:Azacitidine