Src oncogene activates MMP-2 expression via the ERK/Sp1 pathway.
Previous studies demonstrated that activation of Src oncogene increased matrix metalloproteinase-2 (MMP-2) expression in various types of cells. In this study, we elucidated the underlying mechanism of Src-induced MMP-2. We first used murine fibroblast cell line C3H10T1/2 and its v-Src transfectant IV5 to address this issue. RT-PCR and promoter activity assay indicated that Src stimulated MMP-2 via transcriptional activation. Transfection of constitutively active Src into C3H10T1/2 cells also stimulated MMP-2 mRNA expression. Deletion and mutation analysis indicated that the Sp1 site located within the -91/-84 region of human MMP-2 promoter is the major responsive element for Src. Electrophoresis mobility shift assays showed that Src enhanced the binding of Sp1 to this consensus site to stimulate MMP-2 gene expression. We next investigated the signaling pathway that mediated the effect of Src on MMP-2. Our data showed that extracellular signal-regulated kinase (ERK) pathway inhibitor PD98059, but not the c-Jun N-terminal kinase (JNK) inhibitor SP600125, p38 kinase inhibitor SB203580, and PI3K inhibitor wortmannin, attenuated Src-induced MMP-2 promoter activity. These inhibitors did not show significant effect on basal MMP-2 promoter activity in C3H10T1/2 cells. In addition, the dominant negative mutant of ERK-2 suppressed the activation of MMP-2 by Src. Treatment of PD98059 or an Src specific inhibitor PP1 reduced Sp1 DNA binding activity in IV5 cells. Taken together, our results strongly suggest that Src induces MMP-2 expression via transcription activation and the ERK/Sp1 signaling pathway is involved in this process.
Kuo
Lunyu
L
Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan, Republic of China.
Chang
Hui-Chiu
HC
Leu
Tzeng-Horng
TH
Maa
Ming-Chie
MC
Hung
Wen-Chun
WC
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020778:Matrix Metalloproteinase 2 / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D015533:Transcriptional Activation / Q000235:genetics; D015854:Up-Regulation; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*
D005419:Flavonoids; D016329:Sp1 Transcription Factor; D019061:src-Family Kinases; D019950:Mitogen-Activated Protein Kinase 1; D020778:Matrix Metalloproteinase 2; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one