title: 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.
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 (5-25μM) or TA (25-100μM) or combination of Cur (7.5μM) and TA (50μM). Cell viability was measured at 24-72h 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.
authors.affiliation: Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107; Pre-clinical Services, University of North Texas Health Science Center, Fort Worth, TX, 76107. Electronic address: Riyaz.Basha@unthsc.edu.
authors.forename: Riyaz
authors.initials: R
authors.lastname: Basha
authors.affiliation: MD Anderson Cancer Center Orlando, Orlando, FL, 32806.
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Connelly
authors.affiliation: Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107.
authors.forename: Umesh T
authors.initials: UT
authors.lastname: Sankpal
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University, Atlanta, GA, 30322.
authors.forename: Ganji Purnachandra
authors.initials: GP
authors.lastname: Nagaraju
authors.affiliation: Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107.
authors.forename: Hassaan
authors.initials: H
authors.lastname: Patel
authors.affiliation: Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107.
authors.forename: Jamboor K
authors.initials: JK
authors.lastname: Vishwanatha
authors.affiliation: Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107.
authors.forename: Sagar
authors.initials: S
authors.lastname: Shelake
authors.affiliation: Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107.
authors.forename: Leslie
authors.initials: L
authors.lastname: Tabor-Simecka
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University, Atlanta, GA, 30322.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Shoji
authors.affiliation: Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107; Pre-clinical Services, University of North Texas Health Science Center, Fort Worth, TX, 76107; College of Pharmacy, University of North Texas Health Science Center, Fort Worth, TX, 76107.
authors.forename: Jerry W
authors.initials: JW
authors.lastname: Simecka
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University, Atlanta, GA, 30322.
authors.forename: Bassel
authors.initials: B
authors.lastname: El-Rayes
mesh_terms: D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D003474:Curcumin / Q000008:administration & dosage*; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000473:pathology*; D021381:Protein Transport; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D062367:ortho-Aminobenzoates / Q000008:administration & dosage*
chemical_list: D016328:NF-kappa B; D016329:Sp1 Transcription Factor; D062367:ortho-Aminobenzoates; C009500:tolfenamic acid; D003474:Curcumin
keywords: Combination treatment; Curcumin; Pancreatic cancer; Sp1; Tolfenamic acid