title: Tolfenamic acid-induced alterations in genes and pathways in pancreatic cancer cells.
abstract: Non-steroidal anti-inflammatory drugs (NSAIDs) are being tested extensively for their role in the treatment and prevention of several cancers. Typically NSAIDs exhibit anti-tumor activities via modulation of cyclooxygenase (COX)-dependent mechanisms, however, an anti-cancer NSAID tolfenamic acid (TA) is believed to work through COX-independent pathways. Results from our laboratory and others have demonstrated the anti-cancer activity of TA in various cancer models including pancreatic cancer. TA has been shown to modulate certain cellular processes including, apoptosis, reactive oxygen species and signaling. In this study, molecular profiling was performed to precisely understand the mode of action of TA. Three pancreatic cancer cell lines, L3.6pl, MIA PaCa-2, and Panc1 were treated with TA (50 μM for 48 h) and the changes in gene expression was evaluated using the Affymetrix GeneChip Human Gene ST Array platform. Microarray results were further validated using quantitative PCR for seven genes altered by TA treatment in all three cell lines. Functional analysis of differentially expressed genes (2 fold increase or decrease, p < 0.05) using Ingenuity Pathway Analysis software, revealed that TA treatment predominantly affected the genes involved in cell cycle, cell growth and proliferation, and cell death and survival. Promoter analysis of the differentially expressed genes revealed that they are enriched for Sp1 binding sites, suggesting that Sp1 could be a major contributor in mediating the effect of TA. The gene expression studies identified new targets involved in TA's mode of action, while supporting the hypothesis about the association of Sp1 in TA mediated effects in pancreatic cancer.
authors.affiliation: Texas College of Osteopathic Medicine, University of North Texas Health Science Center, TX, USA.
authors.forename: Umesh T
authors.initials: UT
authors.lastname: Sankpal
authors.affiliation: Department of Health Sciences Research, Mayo Clinic, Jacksonville, FL, USA.
authors.forename: Steve
authors.initials: S
authors.lastname: Goodison
authors.affiliation: Texas College of Osteopathic Medicine, University of North Texas Health Science Center, TX, USA.
authors.forename: Michelle
authors.initials: M
authors.lastname: Jones-Pauley
authors.affiliation: Institute for Molecular Medicine, University of North Texas Health Science Center, TX, USA.
authors.forename: Myrna
authors.initials: M
authors.lastname: Hurtado
authors.affiliation: Institute for Molecular Medicine, University of North Texas Health Science Center, TX, USA.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Texas College of Osteopathic Medicine, University of North Texas Health Science Center, TX, USA.|Institute for Molecular Medicine, University of North Texas Health Science Center, TX, USA.
authors.forename: Riyaz
authors.initials: R
authors.lastname: Basha
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D016000:Cluster Analysis; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology; D011401:Promoter Regions, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects* / Q000235:genetics; D016329:Sp1 Transcription Factor / Q000378:metabolism; D062367:ortho-Aminobenzoates / Q000494:pharmacology*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D016329:Sp1 Transcription Factor; C108209:SP1 protein, human; D062367:ortho-Aminobenzoates; C009500:tolfenamic acid
keywords: Sp1; microarray analysis; pancreatic cancer; tolfenamic acid