Preclinical evaluation of 4-[3,5-bis(2-chlorobenzylidene)-4-oxo-piperidine-1-yl]-4-oxo-2-butenoic acid, in a mouse model of lung cancer xenograft.

Abstract

BACKGROUND AND PURPOSE:

4-[3,5-Bis(2-chlorobenzylidene)-4-oxo-piperidine-1-yl]-4-oxo-2-butenoic acid CLEFMA is a new anti-cancer molecule. Here, we investigated changes in apoptosis and inflammatory markers during CLEFMA-induced tumour suppression.

EXPERIMENTAL APPROACH:

Lung adenocarcinoma H441 and A549, and normal lung fibroblast CCL151 cell lines were used, along with a xenograft model of H441 cells implanted in mice. Tumour tissues were analysed by immunoblotting, immunohistochemistry and/or biochemical assays. The ex vivo results were confirmed by performing selected assays in cultured cells.

KEY RESULTS:

CLEFMA-induced cell death was associated with cleavage of caspases 3/9 and PARP. In vivo, CLEFMA treatment resulted in a dose-dependent suppression of tumour growth and (18) F-fluorodeoxyglucose uptake in tumours, along with a reduction in the expression of the proliferation marker Ki-67. In tumour tissue homogenates, the anti-apoptotic markers (cellular inhibitor of apoptosis protein-1(cIAP1), Bcl-xL, Bcl-2, and survivin) were inhibited and the pro-apoptotic Bax and BID were up-regulated. Further, CLEFMA decreased translocation of phospho-p65-NF-κB into the nucleus. In vitro, it inhibited the DNA-binding and transcriptional activity of NF-κB. It also reduced the expression of COX-2 in tumours and significantly depressed serum TNF-α and IL-6 levels. These effects of CLEFMA were accompanied by a reduced transcription and/or translation of the invasion markers VEGF, MMP9, MMP10, Cyclin D1 and ICAM-1.

CONCLUSIONS AND IMPLICATIONS:

Overall, CLEFMA inhibited growth of lung cancer xenografts and this tumour suppression was associated with NF-κB-regulated anti-inflammatory and anti-metastatic effects.

• Authors: Vivek R Yadav, Kaustuv Sahoo, Vibhudutta Awasthi
• Journal: British journal of pharmacology (Br J Pharmacol) Vol. 170 Issue 7 Pg. 1436-48 (Dec 2013) ISSN: 1476-5381 [Electronic] England
• PMID: 24102070 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: © 2013 The British Pharmacological Society.

Chemical References

• 4-(3,5-bis(2-chlorobenzylidene)-4-oxopiperidine-1-yl)-4-oxo-2-butenoic acid
• Antineoplastic Agents
• Apoptosis Regulatory Proteins
• Benzylidene Compounds
• Inflammation Mediators
• Ki-67 Antigen
• Piperidones
• RELA protein, human
• Transcription Factor RelA
• Fluorodeoxyglucose F18

Topics

• Adenocarcinoma (drug therapy, genetics, immunology, metabolism, pathology)
• Adenocarcinoma of Lung
• Animals
• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects)
• Apoptosis Regulatory Proteins (metabolism)
• Benzylidene Compounds (pharmacology)
• Cell Line, Tumor
• Dose-Response Relationship, Drug
• Fluorodeoxyglucose F18
• Gene Expression Regulation, Neoplastic
• Humans
• Inflammation Mediators (metabolism)
• Ki-67 Antigen (metabolism)
• Lung Neoplasms (drug therapy, genetics, immunology, metabolism, pathology)
• Male
• Mice
• Mice, Nude
• Multimodal Imaging
• Phosphorylation
• Piperidones (pharmacology)
• Positron-Emission Tomography
• Signal Transduction (drug effects)
• Time Factors
• Tomography, X-Ray Computed
• Transcription Factor RelA (genetics, metabolism)
• Transfection
• Tumor Burden (drug effects)
• Xenograft Model Antitumor Assays