title: Anti-leukemic response of a NSAID, tolfenamic acid.
abstract: Tolfenamic acid (TA), a non-steroidal anti-inflammatory drug, is known to inhibit human cancer cells and mouse tumor growth in some cancer models; however, its anti-leukemic response has not been evaluated. TA targets specificity protein (Sp) transcription factors that mediate the expression of several genes associated with cancer including survivin, a key member of inhibitor of apoptosis protein family. Our aim was to test the anti-leukemic efficacy of TA in pre-clinical experiments. The anti-leukemic response of TA was determined using Jurkat and Nalm-6 cell lines. Cells were treated with increasing (25/50/75 μM) concentrations of TA, and cell viability was measured at 24, 48, and 72 h post-treatment. TA showed a steady and consistent decrease in cell viability following a clear dose and time dependent response. Apoptosis and cell cycle analysis was performed using flow cytometry. Results showed a significant increase in the apoptotic fraction (annexin V positive) following TA treatment, while cell cycle phase distribution analysis showed G0/G1 arrest. TA-induced apoptosis was further confirmed by examining the activation of caspase 3/7 and the expression of cleaved PARP. TA modulated the expression of critical candidates associated with the early phases of cell cycle and validated its efficacy in causing G0/G1 arrest. The Western blot results revealed that TA significantly decreases Sp1 and survivin expression. These results demonstrate that the anti-leukemic response of TA occurs potentially through targeting Sp1 and inhibiting survivin and suggest the efficacy of TA as a novel therapeutic agent for leukemia.
authors.affiliation: MD Anderson Cancer Center Orlando, Orlando, FL, 32806, USA, Robert.Sutphin@orlandohealth.com.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Sutphin
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Connelly
authors.forename: Chris M
authors.initials: CM
authors.lastname: Lee
authors.forename: Umesh T
authors.initials: UT
authors.lastname: Sankpal
authors.forename: Don
authors.initials: D
authors.lastname: Eslin
authors.forename: Moeez
authors.initials: M
authors.lastname: Khan
authors.forename: Hima
authors.initials: H
authors.lastname: Pius
authors.forename: Riyaz
authors.initials: R
authors.lastname: Basha
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000096:biosynthesis / Q000187:drug effects; D007938:Leukemia / Q000378:metabolism / Q000473:pathology*; D000077022:Survivin; D062367:ortho-Aminobenzoates / Q000494:pharmacology*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D000970:Antineoplastic Agents; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D000077022:Survivin; D062367:ortho-Aminobenzoates; C009500:tolfenamic acid