Benzyl isothiocyanate-mediated inhibition of histone deacetylase leads to NF-kappaB turnoff in human pancreatic carcinoma cells.

Abstract

NF-kappaB/p65 is constitutively activated in pancreatic cancers, where it plays a critical role in the transcriptional activation of multiple cell survival genes. We have previously shown the apoptosis-inducing effects of benzyl isothiocyanate (BITC) in pancreatic cancer cells. We hypothesized that inhibition of NF-kappaB/p65 could be the mechanism of BITC-induced apoptosis. Therefore, the effect of BITC on NF-kappaB/p65 was evaluated in BxPC-3, Capan-2, and normal HPDE-6 cells by Western blotting, transcriptional and DNA-binding activity, and immunohistochemistry in the xenografted tumors. Our results reveal a remarkable decrease in the phosphorylation of NF-kappaB/p65 at Ser(536) in both BxPC-3 and Capan-2 cells by BITC treatment. The expression of NF-kappaB/p65 was downregulated significantly in BxPC-3 cells, whereas it remained unchanged in Capan-2 cells. BITC treatment caused a significant decrease in NF-kappaB transcriptional and DNA-binding activity in both BxPC-3 and Capan-2 cells. A drastic decrease was observed in the expression and reporter activity of cyclin D1 in both the cell lines. Moreover, BITC also caused a significant decrease in the expression and activity of histone deacetylase (HDAC) 1 and HDAC3 in BxPC-3 and HDAC3 in Capan-2 cells. Overexpression of HDAC1 or HDAC3 abrogated the effects of BITC. BITC treatment did not cause any change in HDAC expression in normal HPDE-6 cells. Immunohistochemical analysis of tumors from BITC-treated mice showed significantly reduced staining for NF-kappaB, cyclin D1, HDAC1, and HDAC3 compared with control. Our results suggest inhibition of HDAC1/HDAC3 by BITC as a plausible mechanism of NF-kappaB inactivation, resulting in the in vitro and in vivo growth suppression of pancreatic cancer cells.

• Authors: Sanjay Batra, Ravi P Sahu, Prabodh K Kandala, Sanjay K Srivastava
• Journal: Molecular cancer therapeutics (Mol Cancer Ther) Vol. 9 Issue 6 Pg. 1596-608 (Jun 2010) ISSN: 1538-8514 [Electronic] United States
• PMID: 20484017 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Cyclin-Dependent Kinase Inhibitor p21
• Histone Deacetylase Inhibitors
• Isothiocyanates
• NF-kappa B
• Neoplasm Proteins
• Transcription Factor RelA
• Cyclin D1
• benzyl isothiocyanate
• DNA
• Histone Acetyltransferases
• Histone Deacetylases

Topics

• Apoptosis (drug effects)
• Cell Line, Tumor
• Cyclin D1 (genetics, metabolism)
• Cyclin-Dependent Kinase Inhibitor p21 (metabolism)
• DNA (metabolism)
• Down-Regulation (drug effects)
• Drug Resistance, Neoplasm (drug effects)
• Gene Expression Regulation, Neoplastic (drug effects)
• Genes, Reporter
• Histone Acetyltransferases (antagonists & inhibitors)
• Histone Deacetylase Inhibitors (pharmacology)
• Histone Deacetylases (metabolism)
• Humans
• Isothiocyanates (pharmacology)
• NF-kappa B (genetics, metabolism)
• Neoplasm Proteins (genetics, metabolism)
• Pancreatic Neoplasms (enzymology, genetics, pathology)
• Promoter Regions, Genetic (genetics)
• Protein Binding (drug effects)
• Transcription Factor RelA (metabolism)
• Transcription, Genetic (drug effects)
• Xenograft Model Antitumor Assays