This Article Full Text Full Text (PDF) Supplementary Data Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kwak, H.-J. Articles by Hong, S.-I. Search for Related Content PubMed PubMed Citation Articles by Kwak, H.-J. Articles by Hong, S.-I.

---

Signalling and Regulation

Transforming Growth Factor-ß1 Induces Tissue Inhibitor of Metalloproteinase-1 Expression via Activation of Extracellular Signal-Regulated Kinase and Sp1 in Human Fibrosarcoma Cells

¹ Laboratory of Functional Genomics, ² Department of Neurosurgery, and ³ Laboratory Medicine and Clinical Pathology, Korea Institute of Radiological and Medical Sciences, Seoul, Korea and ⁴ Research Institute and hospital, National Cancer Center, Goyang, Korea

Requests for reprints: Seok-Il Hong, Laboratory of Functional Genomics, Korea Institute of Radiological and Medical Sciences, 215-4 Nowon-Ku, Gongneung-Dong, Seoul 139-706, Korea. Phone: 82-2-970-1260; Fax: 82-2-970-2402. E-mail: hongsicp{at}kcch.re.kr

The net balance of matrix metalloproteinases (MMP) and tissue inhibitor of metalloproteinases (TIMP) system has been known to be a key factor in tumor cell invasion. In the present study, we investigated the molecular mechanisms of anti-invasive and antimigrative activity of transforming growth factor (TGF)-ß1 on HT1080 human fibrosarcoma cells. In in vitro Matrigel invasion and Transwell migration assays, TGF-ß1 dose-dependently inhibited the invasion and migration of HT1080 cells, respectively. Gelatin zymography, Western blot, and real-time PCR analysis showed that TGF-ß1 enhanced the expression and secretion of MMP-2, TIMP-1, and, to a lesser degree, MMP-9 but not membrane type 1-MMP and TIMP-2. The addition of recombinant TIMP-1 protein reduced the Matrigel invasion and Transwell migration of HT1080 cells, similar to TGF-ß1. Because augmentation of TIMP-1 might be the major factor for the anti-invasive and antimigrative activity of TGF-ß1, we investigated possible molecular mechanisms responsible for the expression of TIMP-1 induced by TGF-ß1. Treatment of HT1080 cells with TGF-ß1 rapidly phosphorylated three mitogen-activated protein kinases [MAPK; extracellular signal-regulated kinase 1/2 (ERK1/2), p38, and c-Jun NH₂-terminal kinase] and Akt. Among these kinases, the inhibition of only ERK1/2 pathway by PD98059, a specific inhibitor of MAPK/ERK kinase(MEK)-1, and transfection of dominant-negative MEK 1 effectively blocked the TIMP-1 induction by TGF-ß1. Mithramycin, a specific inhibitor of Sp1 transcription factor, but not curcumin, an inhibitor of activator protein-1, and transfection of Sp1 small interfering RNA significantly inhibited the TGF-ß1-induced expression of TIMP-1. In addition, electrophoretic mobility shift assay showed that TGF-ß1 up-regulated Sp1 DNA-binding activity, and PD98059 and mithramycin effectively inhibited these events. Finally, pretreatment of HT1080 cells with PD98059 and mithramycin, but not curcumin, restored the invasive activity of these cells. Taken together, these data suggest that TGF-ß1 modulates the net balance of the MMPs/TIMPs the systems in HT1080 cells for anti-invasion and antimigration by augmenting TIMP-1 through ERK1/2 pathway and Sp1 transcription factor. (Mol Cancer Res 2006;4(3):209–20)

This article has been cited by other articles:

				A. V. Miller, S. E. Alvarez, S. Spiegel, and D. A. Lebman Sphingosine Kinases and Sphingosine-1-Phosphate Are Critical for Transforming Growth Factor {beta}-Induced Extracellular Signal-Regulated Kinase 1 and 2 Activation and Promotion of Migration and Invasion of Esophageal Cancer Cells Mol. Cell. Biol., June 15, 2008; 28(12): 4142 - 4151. [Abstract] [Full Text] [PDF]

				F. Barcelo, C. Scotta, M. Ortiz-Lombardia, C. Mendez, J. A. Salas, and J. Portugal Entropically-driven binding of mithramycin in the minor groove of C/G-rich DNA sequences Nucleic Acids Res., April 1, 2007; 35(7): 2215 - 2226. [Abstract] [Full Text] [PDF]

				P.-P. Kuang, X.-H. Zhang, C. B. Rich, J. A. Foster, M. Subramanian, and R. H. Goldstein Activation of elastin transcription by transforming growth factor-beta in human lung fibroblasts Am J Physiol Lung Cell Mol Physiol, April 1, 2007; 292(4): L944 - L952. [Abstract] [Full Text] [PDF]