KEGG analysis of differentially expressed genes induced by CBHA and TSA To extend the in silico examination of the differen tially regulated genes by IPA, table 1 we subjected DEGs that were common to TSA and CBHA to KEGG analysis. The KEGG program is designed to convert the mo lecular interactions and gene networks into biologic ally functional pathways. The KEGG analysis revealed that CBHA and TSA elicited a number of overlapping pathways, regardless of the duration of the treatment. Thus, phosphatidylinositol metabolism and signaling and MAPK pathways were preeminent in H9c2 cells exposed to either TSA or CBHA at 6h. Furthermore, the putative PTEN PI3K AKT/PKB signaling pathways were connected with numerous genes involved in the metabolism of pyru vate, citrate and amino acids, as well as in the inter mediary metabolism of purines and pyrimidines.
The emergence of gene networks known to regulate cell cycle and DNA replication, metabolism of xenobiotics, oxidative stress and extracellular matrix were also common in H9c2 cells incubated with either CBHA Inhibitors,Modulators,Libraries or TSA for 24h. Based on these observations we surmise that similar HDACI induced gene networks were uncovered Inhibitors,Modulators,Libraries by IPA and KEGG analyses. A putative involvement of MAPK pathways in the action of pan HDAC inhibitors The network analyses of genes that were differentially regulated by CBHA and TSA, regardless of whether it was done by IPA or KEGG programs, strongly predicted a role of PTEN PI3K AKT/PKB and MAPK signaling pathways in the actions of HDACIs.
We reported earlier that both CBHA and TSA potently induced the expres sion of Inhibitors,Modulators,Libraries PTEN and concomitant reduction in PI3K and AKT phosphorylation in H9c2 cells as well as in the in tact heart. To test a potential role of MAP Inhibitors,Modulators,Libraries kinases, we extracted proteins from H9c2 cells incubated with CBHA or TSA for various time intervals and assessed the steady state levels of total and phosphorylated ERK, Inhibitors,Modulators,Libraries JNK and p38 MAPK. As shown in Figure 11, an expos ure to TSA for 4h led to a reduced phosphorylation of ERK and its phosphorylation remained inhibited until 24h. TSA treatment also significantly suppressed phosphorylation of p38 as early as 2h. Finally, an exposure of H9c2 cells to CBHA resulted in a reduction of pERK at 4h, while the levels of p p38 kinase were not significantly affected by CBHA. The temporal changes in the regulation of JNK in response to CBHA or TSA were inconclusive.
Finally, it should be noted that neither TSA nor CBHA altered the steady state levels of total ERK or p38 kinases. Frequency of putative transcription factor binding sites in differentially expressed genes in response to CBHA and TSA www.selleckchem.com/products/MG132.html With an aim to elucidate potentially common pathways involved in the induction of genes by CBHA and TSA, we extended gene network analyses by an in silico exam ination of transcription factor binding sites in the promoters of DEGs.