Comparative Proteome Profiling and Functional Analysis of Chronic Myelogenous Leukemia Cell Lines

Abstract

The aim of the present study was the molecular profiling of different Ph+ chronic myelogenous leukemia (CML) cell lines (LAMA84, K562, and KCL22) by a proteomic approach. By employing two-dimensional gel electrophoresis combined with mass spectrometry analysis, we have identified 191 protein spots corresponding to 142 different proteins. Among these, 63% were cancer-related proteins and 74% were described for the first time in leukemia cells. Multivariate analysis highlighted significant differences in the global proteomic profile of the three CML cell lines. In particular, the detailed analysis of 35 differentially expressed proteins revealed that LAMA84 cells preferentially expressed proteins associated with an invasive behavior, while K562 and KCL22 cells preferentially expressed proteins involved in drug resistance. These data demonstrate that these CML cell lines, although representing the same pathological phenotype, show characteristics in their protein expression profile that suggest different phenotypic leukemia subclasses. These data contribute a new potential characterization of the CML phenotype and may help to understand interpatient variability in the progression of disease and in the efficacy of a treatment.

Chronic Myelogenous Leukemia: Approaches to Pharmacological Resistance

“…After a comparative functional and proteomic analysis of CML cell lines [5], we worked many years trying to overcome IM-resistance with new compounds and we studied the effects of Carboxyamidotriazole (CAI) and later of Carboxyamidotriazole Orotate (CTO) on CML cell lines IM-resistant. Calcium is a well-known second messenger involved in regulation of cell proliferation and apoptosis.…”

Production of an egg yolk antibody against Parietaria judaica 2 allergen

“…A 2-dimensional gel electrophoresis (2DE) PAGE was performed as described previously (Fontana et al, 2008). Briefly, proteins were solubilized in a buffer containing 8 M urea, 4% wt/vol 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate, 40 mM Tris, and 65 mM dithioerythritol (DTE), and protein concentrations were determined using the Coomassie Protein Assay (Pierce).…”

Proteomic analysis of Parietaria judaica pollen and allergen profiling by an immunoproteomic approach

“…The first dimension, equilibration steps and the second dimension were performed as previously described (Fontana et al 2007). …”