Proteomics approach in classifying the biochemical basis of the anticancer activity of the new olomoucine-derived synthetic cyclin-dependent kinase inhibitor, bohemine

Abstract

The aim of this study was to use two-dimensional electrophoresis (2-DE) coupled with multivariate principal component analysis (PCA) to characterize the quantitative changes in the protein composition of the CEM T-lymphoblastic leukemia cell line after treatment with bohemine (BOH), a synthetic olomoucin-derived cyclin-dependent kinase inhibitor (CDKI). Cell classification, reflecting protein patterns, clearly distinguished two main groups: one group consists of 9, 12 and 24 h treated BOH cells while the second is represented by the 0 and 24 h control untreated cells and the 6 h BOH-exposed CEM lymphoblasts. Discriminant protein spots differentially expressed in the BOH-treated CEM cells were selected for identification by matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS) or electrospray ionization-tandem MS (ESI-MS/MS). Five of the selected protein spots were unequivocally identified as alpha-enolase, triosephosphate isomerase, eukaryotic initiation factor 5A, and alpha- and beta-subunits of Rho GDP-dissociation inhibitor 1. These proteins, all significantly downregulated in CEM T-lymphoblast leukemia in the course of BOH treatment, are known to play an important role in cellular functions such as glycolysis, protein biosynthesis, and cytoskeleton rearrangement. These results indicate that the cellular effects of olomoucine-derived CDKIs are not dependent on their ability to inhibit CDKs and could be mediated by several factors such as a decrease in protein synthesis and/or glycolysis which in turn diminishes the ability of cancer cells to function.

Apoptotic effect of resveratrol on human T‐ALL cell line CCRF‐CEM is unlikely exerted through alteration of BAX and BCL2 promoter methylation

“…It has been demonstrated that red polyphenols which also contain resveratrol have a role in downregulating ubiquitin-like, containing PHD and RING finger domains 1 as an epigenetic equilibrium which regulates both DNA methylation and histone modifications. 28,29 There are evidence which clarify the correlation of resveratrol effect in the BAX and BCL2 genes in molecular levels, 30,31 although the mechanisms behind the gene regulation are yet to be comprehensively described.…”

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“…T-lymphoblastic cell line CEM was treated by bohemine and the changes of the protein composition, as visualized in two-dimensional electrophoresis, were quantitated. In bohemine treated cells, five proteins were found to be downregulated, namely α-enolase, triosephosphate isomerase, initiation factor 5A, and α-and β-subunits of Rho GDP-dissociation inhibitor 1 (Kováȓová et al, 2000). These proteins play significant roles in glycolysis, proteosynthesis, and in cytoskeleton rearrangement.…”