Inhibition of Histone Deacetylase Activity Promotes Invasion of Human Cancer Cells through Activation of Urokinase Plasminogen Activator*
- Departments of ‡Cancer Biology and Pharmacology and §Neurosurgery, University of Illinois College of Medicine, Peoria, Illinois 61656
- 1 To whom correspondence should be addressed: Program of Cancer Biology, University of Illinois College of Medicine, One Illini Drive, Peoria, IL 61605. Tel.: 309-671-3445; Fax: 309-671-3442; E-mail: jsrao{at}uic.edu.
Abstract
Histone acetylation plays an important role in chromatin remodeling and gene expression. The molecular mechanisms involved in differential regulation of urokinase plasminogen activator (uPA) gene expression are not fully understood. In this study, we investigated whether histone deacetylation was involved in repression of uPA expression in human cancer cells. Induction of uPA expression by histone deacetylase (HDAC) inhibitors trichostatin A (TSA), sodium butyrate, and scriptaid was observed in all three different types of human cancer cells examined. Chromatin immunoprecipitation assays showed that the induction of uPA expression by TSA was accompanied by a remarkable increase of acetylation of histones H3 and H4, which are associated with the uPA promoter region in human cancer cells. These results were further substantiated by the findings of a restriction enzyme accessibility assay and TSA-stimulated uPA promoter activity through the inhibition of HDAC activity. In vitro Matrigel invasion assays showed that induction of uPA expression by HDAC inhibitors in human cancer cells resulted in a significant increase of cancer cell invasion. Furthermore, HDAC1 knockdown by small interference RNA stimulated uPA expression and cancer cell invasion. In conclusion, this study demonstrates the important role of histone modifications in regulating uPA gene expression and raises a possibility that the use of HDAC inhibitors in patients as cancer therapy may paradoxically establish metastasis through up-regulation or reactivation of uPA.
Footnotes
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↵2 The abbreviations used are: uPA, urokinase plasminogen activator; HDAC, histone deacetylase; HDAI, histone deacetylase inhibitor; TSA, trichostatin A; NaB, sodium butyrate; SCR, scriptaid; 5-aza, 5-aza-2′-deoxycytidine; siRNA, small interfering RNA; shRNA, small hairpin RNA; RNAi, RNA interference; ChIP, chromatin immunoprecipitation; RT, reverse transcription; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; CMV, cytomegalovirus.
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↵* This work was supported by NCI, National Institutes of Health (NIH) Grants CA75557, CA92393, CA95058, and CA116708, by NINDS, NIH Grants NS47699 and NS057529, and by Caterpillar, Inc., OSF Saint Francis, Inc., Peoria, IL (to J. S. R.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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- Received July 17, 2007.
- Revision received September 27, 2007.
