Protein fingerprints of anti-cancer effects of cyclin-dependent kinase inhibition: identification of candidate biomarkers using 2-D liquid phase separation coupled to mass spectrometry.
The purpose of this study was to apply a recently introduced proteomic based approach to identify candidate biomarkers of the response to anticancer activity of cyclin-dependent kinase inhibitor, bohemine. Mapping of the total protein expression of CEM lymphoblastic leukemia cells following bohemine treatment was performed by 2-D liquid phase separation. Proteins were fractionated by isoelectric points in pH gradient in the first dimension and each of these pI protein fractions was further separated by hydrophobicity using non-porous silica reverse phase chromatography in the second dimension. 2-D protein expression maps of control untreated and bohemine treated cells were generated and inter-sample comparison was performed. Most of the differentially expressed proteins were present at a decreased level after bohemine treatment while there were four proteins, which were up regulated. These proteins representing candidate biomarkers of cancer cell response to the treatment were selected for identification by mass spectrometry. Our results demonstrating down regulation of three histone variants, different in their pI and hydrophobicity, in response to bohemine indicated that anti-mitotic and anti-cancer activities of this compound may be associated with epigenetic regulation at the level of chromatin structure. Furthermore, crk-like adaptor scaffolding protein represents a new important protein family affected by bohemine. This strategy is valuable for comprehensive proteomic analysis of cellular protein targets and pathways that are relevant to anticancer activity of cyclin-dependent kinase inhibition.
Skalnikova
Helena
H
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Rumburska 89, 27721 Libechov, Czech Republic.
Halada
Petr
P
Dzubak
Petr
P
Hajduch
Marian
M
Kovarova
Hana
H
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D015415:Biomarkers / Q000737:chemistry*; D014408:Biomarkers, Tumor / Q000235:genetics*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002843:Chromatin / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D015180:Electrophoresis, Gel, Two-Dimensional; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D013058:Mass Spectrometry / Q000379:methods*; D008956:Models, Chemical; D009369:Neoplasms / Q000378:metabolism*; D010449:Peptide Mapping; D020543:Proteome; D040901:Proteomics / Q000379:methods; D011687:Purines / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000970:Antineoplastic Agents; D015415:Biomarkers; D014408:Biomarkers, Tumor; D002843:Chromatin; D020543:Proteome; D011687:Purines; C118034:bohemine; D018844:Cyclin-Dependent Kinases