Fibroblast growth factor-1 transcriptionally induces membrane type-1 matrix metalloproteinase expression in prostate carcinoma cell line.
BACKGROUND
We and others have shown that the matrix metalloproteinases, MT1-MMP is overexpressed in human prostate PIN lesions and invasive cancers compared to normal prostate epithelium. However, the mechanism for this overexpression is not understood. Evidence from our laboratory and others has indicated that fibroblast growth factors (FGFs) can regulate the expression of certain matrix metalloproteinase. In addition, human prostate fibroblasts are known to express certain FGFs, including FGF-1. The purpose of the work in this paper was to determine the mechanism involved in FGF-1 induced MT1-MMP expression in prostate carcinoma cells.


METHODS
We tested the ability of recombinant FGF-1 to induce MT1-MMP expression in prostate carcinoma cell line, LNCaP cells. We measured the MT1-MMP message by using Northern analyses and protein levels by Western analysis after FGF-1 treatment. Downstream signaling was investigated using dominant negative constructs for FGFR-1 and signal transducer and activator of transcription-3 (STAT3). Transient transfection was performed using reporter plasmids of the MT1-MMP gene promoter region (7.2 kb) linked to the firefly luciferase gene in the pGL3-Basic vector. For dominant negative studies FGFR-1 dominant negative plasmid in PCEP4 vector or STAT3 dominant negative plasmid in pCMV-1 vector was co-transfected with the MT1-MMP reporter plasmid.


RESULTS
Recombinant FGF-1 significantly induced MT1-MMP expression in LNCaP prostate carcinoma cells. MT1-MMP message increased with FGF-1 treatment compared to that of untreated control LNCaP cells. Quantitation by digital image analysis revealed that this increase was twofold over untreated LNCaP cells. Treatment of pGL3-MT1-MMP-luciferase transfected cells with FGF-1 resulted in a twofold to fourfold increase in luciferase enzyme activity compared with untreated cells. Co-transfection of LNCaP with human MT1-MMP reporter construct and a dominant negative FGFR1 mutant showed that FGF-1-induced MT1-MMP expression in LNCaP cells was completely inhibited by the mutated FGFR-1, indicating that FGF receptor (FGFR) activation is necessary for induction of MT1-MMP. Further, expression of dominant negative STAT3 inhibited the FGF-1-induced transactivation of the human MT1-MMP 7.2-kb promoter.


CONCLUSIONS
From these data, we conclude that FGF-1 induces MT1-MMP expression in prostate carcinoma cells through a transcriptional mechanism mediated through the FGFR and the transcription factor, STAT3. These results confirm earlier data indicating that acidic FGF and STAT3 are involved in the signaling leading to the expression of a MMP. Our findings support the idea that paracrine and autocrine factors play an important role in the regulation of MT1-MMP in human prostate carcinoma cells.
Udayakumar
T S
TS
Department of Cell Biology and Anatomy, University of Arizona Health Sciences Center, Tucson, Arizona 85724, USA.
Nagle
Raymond B
RB
Bowden
G Tim
GT
D015152:Blotting, Northern; D015153:Blotting, Western; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000378:metabolism; D004790:Enzyme Induction; D016220:Fibroblast Growth Factor 1 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics / Q000502:physiology; D006801:Humans; D008297:Male; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000235:genetics; D012334:RNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics; D051496:Receptor, Fibroblast Growth Factor, Type 1; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D050796:STAT3 Transcription Factor; D015534:Trans-Activators / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics / Q000502:physiology; D014162:Transfection
D004268:DNA-Binding Proteins; D012334:RNA, Neoplasm; D017468:Receptors, Fibroblast Growth Factor; D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D015534:Trans-Activators; D016220:Fibroblast Growth Factor 1; C496348:FGFR1 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases