Modulation of angiogenesis-related proteins synthesis by sodium butyrate in colon cancer cell line HT29

Abstract

Sodium butyrate (NaB), a short-chain fatty acid naturally present in the human colon, is able to induce cell cycle arrest, differentiation and apoptosis in colon cancer cells. In addition to these effects, we investigated the effect of NaB on two angiogenesis-related proteins in a colon carcinoma cell line (HT29): vascular endothelial growth factor (VEGF), the most potent angiogenic factor, and hypoxia-inducible factor (HIF)-1alpha, the main transcription activator of the VEGF gene, which are both constitutively expressed at high levels in HT29 also in normoxic conditions. NaB treatment had a different effect on VEGF165 and HIF-1alpha expression. In fact, it induced a dose-dependent down regulation of the VEGF165 protein level that was not paralleled by a concomitant down regulation of the corresponding mRNA, suggesting a post-translational regulation of the factor. Conversely, after 24 h of treatment all the tested NaB concentrations reduced the HIF-1alpha protein level, whereas after a longer time of exposure HIF-1alpha level increased in the presence of a high NaB concentration (2 mM) with a concomitant increase in HIF-1alpha mRNA. These results indicate that NaB, besides regulating other fundamental cellular processes, is able to modulate the expression of two important angiogenesis-related molecules and suggested a further possible clinical application of this short-chain fatty acid as an anti-angiogenic compound in association with conventional chemotherapeutic agents.

Butyrate-induced proapoptotic and antiangiogenic pathways in EAT cells require activation of CAD and downregulation of VEGF

“…Similarly, studies have also shown to inhibit the translocation of HIF-1a transcription in response to BuA-treatment [32]. We and others have previously shown the direct effect of BuA on VEGF and its receptors [31,28,29]. Consistent with the previous reports, our data demonstrate a dose-dependent inhibition of VEGF secretion in vivo.…”

“…The ability of BuA to modulate NF-jB activity may arise from its ability to inhibit histone deacetylases. The antiproliferative effect of BuA on transformed cells is well established, and it has emerged as an antiangiogenic agent by directly repressing the expression of angiogenic ligands or indirectly interfering with endothelial cell proliferation/enzymes required for angiogenesis [28][29][30]. We have previously shown that BuA could repress gene expression of VEGF/KDR systems in Ehrlich ascites tumor (EAT) cells in vitro [31].…”

Butyrate down-regulates CD44 transcription and liver colonisation in a highly metastatic human colon carcinoma cell line

“…In addition, a recent publication by Pelizzaro et al (2002), show that butyrate induced a dose-dependent down-regulation of VEGF165 protein, a potent angiogenic factor and vis-à-vis up-regulates the hypoxia-inducible factor (HIF)-1a. Our reported effects on CD44 and the effects reported by our group previously (Barshishat et al, 2000) and by Pelizarro et al (2002) may induce altogether the significant effects induced by butyrate observed in vivo on metastatic formation. Nevertheless, the pan-deregulatory effect exerted by butyrate in the entire CD44 gene seems to play a key role in the complete abolishment of metastatic ability exhibited by HM7 cells.…”

Isolation and In vitro Propagation of Tumorigenic Breast Cancer Cells with Stem/Progenitor Cell Properties

“…Total mRNA was extracted by means of Trizol (Life Technologies, Frederick, MD) according to the instructions of the manufacturer. Reverse transcription-PCR amplification of VEGF-A and VEGF-C mRNA was carried out using the GeneAmp Gold RNA PCR reagent kit (Perkin-Elmer Biosystems, Foster City, CA) and the human VEGF-C PCR Primer Pair kit, respectively; samples were processed as described elsewhere (7). h-Actin was used as housekeeping gene and results were expressed as normalized densitometric units.…”