Small molecule tolfenamic acid and dietary spice curcumin treatment enhances antiproliferative effect in pancreatic cancer cells via suppressing Sp1, disrupting NF-kB translocation to nucleus and cell cycle phase distribution

• Type of Material: Article
• Authors: Riyaz Basha, University of North Texas Health Science Center; Sarah F. Connelly, MD Anderson Cancer Center Orlando; Umesh T. Sankpal, University of North Texas Health Science Center; Purnachandra Ganji, Emory University; Hassaan Patel, University of North Texas Health Science Center; Jamboor K. Vishwanatha, University of North Texas Health Science Center; Sagar Shelake, University of North Texas Health Science Center; Leslie Tabor-Simecka, University of North Texas Health Science Center; Mamoru Shoji, Emory University; Jerry W. Simecka, University of North Texas Health Science Center; Bassel El-Rayes, Emory University
• Language: English
• Date: 2016-05-01
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2016 Elsevier Inc.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jnutbio.2016.01.003
• Title of Journal or Parent Work: Journal of Nutritional Biochemistry
• ISSN: 0955-2863
• Volume: 31
• Start Page: 77
• End Page: 87
• Grant/Funding Information: This work was partially supported by a grant from the Shirley E. Noland Foundation (awarded to RB), Institute for Cancer Research and Pre-clinical Services, UNTHSC (RB & JS).; JKV is supported by a grant (1P20 MD006882) from the National Institute on Minority Health and Health Disparities, NIH.
• Keywords: APOPTOSIS; ENDOTHELIAL GROWTH-FACTOR; AGENTS; Pancreatic cancer; GASTRIC-CANCER; Science & Technology; Curcumin; Life Sciences & Biomedicine; Tolfenamic acid; TRANSCRIPTION FACTOR SP1; Combination treatment; PREVENTION; FACTOR EXPRESSION; SURVIVIN; Sp1; Nutrition & Dietetics; MEDIATED ACTIVATION; BREAST-CANCER; Biochemistry & Molecular Biology
• Research Categories: Health Sciences, Immunology; Biology, Molecular

Abstract

Combination of dietary/herbal spice curcumin (Cur) and COX inhibitors has been tested for improving therapeutic efficacy in pancreatic cancer (PC). The objective of this study was to identify agent with low toxicity and COX-independent mechanism to induce PC cell growth inhibition when used along with Cur. Anticancer NSAID, tolfenamic acid (TA) and Cur combination were evaluated using PC cell lines. L3.6pl and MIA PaCa-2 cells were treated with Cur (5-25 μM) or TA (25-100 μM) or combination of Cur (7.5 μM) and TA (50 μM). Cell viability was measured at 24-72 h posttreatment using CellTiter-Glo kit. While both agents showed a steady/consistent effect, Cur + TA caused higher growth inhibition. Antiproliferative effect was compared with COX inhibitors, Ibuprofen and Celebrex. Cardiotoxicity was assessed using cordiomyocytes (H9C2). The expression of Sp proteins, survivin and apoptotic markers (western blot), caspase 3/7 (caspase-Glo kit), Annexin-V staining (flow cytometry), reactive oxygen species (ROS) and cell cycle phase distribution (flow cytometry) was measured. Cells were treated with TNF-α, and NF-kB translocation from cytoplasm to nucleus was evaluated (immunofluorescence). When compared to individual agents, combination of Cur + TA caused significant increase in apoptotic markers, ROS levels and inhibited NF-kB translocation to nucleus. TA caused cell cycle arrest in G0/G1, and the combination treatment showed mostly DNA synthesis phase arrest. These results suggest that combination of Cur + TA is less toxic and effectively enhance the therapeutic efficacy in PC cells via COX-independent mechanisms.