Cheng-Wei Lin and Shing-Chuan Shen contributed equally to the present work.
12-O-tetradecanoylphorbol-13-acetate-induced invasion/migration of glioblastoma cells through activating PKCα/ERK/NF-κB-dependent MMP-9 expression†
Article first published online: 10 MAY 2010
Copyright © 2010 Wiley-Liss, Inc.
Journal of Cellular Physiology
Volume 225, Issue 2, pages 472–481, November 2010
How to Cite
Lin, C.-W., Shen, S.-C., Chien, C.-C., Yang, L.-Y., Shia, L.-T. and Chen, Y.-C. (2010), 12-O-tetradecanoylphorbol-13-acetate-induced invasion/migration of glioblastoma cells through activating PKCα/ERK/NF-κB-dependent MMP-9 expression. J. Cell. Physiol., 225: 472–481. doi: 10.1002/jcp.22226
- Issue published online: 23 AUG 2010
- Article first published online: 10 MAY 2010
- Manuscript Accepted: 16 APR 2010
- Manuscript Received: 26 NOV 2009
- National Science Council of Taiwan. Grant Numbers: NSC96-2320-B-038-031-MY3, 98-2320-B-038-002-MY3
- Taipei Medical University—Shuang Ho Hospital. Grant Number: 98TMU-SHH-01-2
An increase in MMP-9 gene expression and enzyme activity with stimulating the migration of GBM8401 glioma cells via wound healing assay by 12-O-tetradecanoylphorbol-13-acetate (TPA) was detected in glioblastoma cells GBM8401. TPA-induced translocation of protein kinase C (PKC)α from the cytosol to membranes, and migration of GBM8401 elicited by TPA was suppressed by adding the PKCα inhibitors, GF109203X and H7. Activation of extracellular signal-regulated kinase (ERK) and c-Jun-N-terminal kinase (JNK) by TPA was identified, and TPA-induced migration and MMP-9 activity was significantly blocked by ERK inhibitor PD98059 and U0126, but not JNK inhibitor SP600125. Activation of NF-κB protein p65 nuclear translocation and IκBα protein phosphorylation with increased NF-κB-directed luciferase activity by TPA were observed, and these were blocked by the PD98059 and IkB inhibitor BAY117082 accompanied by reducing migration and MMP-9 activity induced by TPA in GBM8401 cells. Transfection of GBM8401 cells with PKCα siRNA specifically reduced PKCα protein expression with blocking TPA-induced MMP-9 activation and migration. Additionally, suppression of TPA-induced PKCα/ERK/NK-κB activation, migration, and MMP-9 activation by flavonoids including kaempferol (Kae; 3,5,7,4′-tetrahydroxyflavone), luteolin (Lut; 5,7,3′4′-tetrahydroxyflavone), and wogonin (Wog; 5,7-dihydroxy-8-methoxyflavone) was demonstrated, and structure–activity relationship (SAR) studies showed that hydroxyl (OH) groups at C4′ and C8 are critical for flavonoids' action against MMP-9 enzyme activation and migration/invasion of glioblastoma cells elicited by TPA. Application of flavonoids to prevent the migration/invasion of glioblastoma cells through blocking PKCα/ERK/NF-κB activation is first demonstrated herein. J. Cell. Physiol. 225: 472–481, 2010. © 2010 Wiley-Liss, Inc.