title: Activating transcription factor 2 (ATF2) controls tolfenamic acid-induced ATF3 expression via MAP kinase pathways.
abstract: Tolfenamic acid (TA) is a non-steroidal anti-inflammatory drug associated with anti-tumorigenic and pro-apoptotic properties in animal and in vitro models of cancer. However, the underlying cellular mechanisms by which TA exerts its effects are only partially understood. Activating transcription factor 3 (ATF3) is a member of the ATF/CREB subfamily of the basic region-leucine zipper family and has been known as a tumor suppressor in human colorectal cancer cells. The present study was performed to observe whether ATF3 mediates TA-induced apoptosis and to elucidate the molecular mechanism of ATF3 transcription induced by TA. TA treatment and ectopic expression of ATF3 increased apoptosis, whereas knockdown of ATF3 resulted in significant repression of TA-activated apoptosis. The TA treatment also induced ATF3 promoter activity. Internal deletion and point mutation of the predicted ATF/C/EBP binding site in ATF3 promoter abolished luciferase activation by TA. Overexpression of ATF2 resulted in significant increase in ATF3 promoter activity, and electrophoretic mobility shift assay identified this region as a core sequence to which ATF2 binds. TA treatment resulted in an increase in ATF2 phosphorylation, which was followed by a subsequent increase in ATF3 transcription. Knock down of ATF2 abolished TA-induced ATF3 expression. We further provide evidence that TA leads to increases in phospho-p38 MAPK, JNK and ERK levels. Inhibition of these pathways using selective inhibitors and dominant negative constructs ameliorated TA-induced ATF3 expression and promoter activities. The current study shows that TA stimulates ATF3 expression and subsequently induces apoptosis. These pathways are mediated through phosphorylation of ATF2, which is mediated by p38 MAPK-, JNK- and ERK-dependent pathways.
authors.affiliation: Department of Pathobiology, Laboratory of Environmental Carcinogenesis, College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA.
authors.forename: S-H
authors.initials: SH
authors.lastname: Lee
authors.forename: J H
authors.initials: JH
authors.lastname: Bahn
authors.forename: N C
authors.initials: NC
authors.lastname: Whitlock
authors.forename: S J
authors.initials: SJ
authors.lastname: Baek
mesh_terms: D051698:Activating Transcription Factor 2 / Q000235:genetics / Q000502:physiology*; D051700:Activating Transcription Factor 3 / Q000235:genetics*; D001483:Base Sequence; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000473:pathology; D004247:DNA; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D020935:MAP Kinase Signaling System; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D062367:ortho-Aminobenzoates / Q000494:pharmacology*
chemical_list: C497105:ATF2 protein, human; C490449:ATF3 protein, human; D051698:Activating Transcription Factor 2; D051700:Activating Transcription Factor 3; D062367:ortho-Aminobenzoates; C009500:tolfenamic acid; D004247:DNA