Expression of matrix metalloproteinase-2 and -9 and their inhibitors, tissue inhibitor of metalloproteinase-1 and -2, in primary cultures of human prostatic stromal and epithelial cells.
The production of matrix metalloproteinases (MMPs) by prostatic epithelial and/or neighboring stromal cells is considered to be a property that gives cells the capability to penetrate extracellular matrix barriers in normal or neoplastic growth. In order to examine the role of MMPs in the prostate, we evaluated the expression of MMP-2 and -9 and the tissue inhibitors of matrix metalloproteinases (TIMP)-1 and -2 in primary cultures of prostatic stromal and epithelial cells. These cells were isolated from normal tissues of the different zones of the prostate, from benign prostatic hyperplasia (BPH) and from cancer. Stromal cells, regardless of tissue of origin, secreted the 72-kDa proenzyme form of MMP-2, whereas conditioned media (CM) from epithelial cells demonstrated little/no pro-MMP-2 as examined by zymography. Either type of cell did not secrete MMP-9. RT-PCR evaluation showed stromal cells expressed transcripts for MMP-2, but not for MMP-9. Transcripts for MMP-9 were detected in epithelial cells, although no MMP-9 activity was detected in their CM. Treatment of stromal cells with 1 or 10 ng/ml of transforming growth factor-beta (TGF-beta) moderately increased secretion of pro-MMP-2 protein with little change in MMP-2 RNA. However, treatment of epithelial cells with TGF-beta induced expression and secretion of both MMP-2 and-9. The effect of TGF-beta on expression of MMPs by epithelial cells was not duplicated or affected by treatment with insulin-like growth factor (IGF)-1, epidermal growth factor (EGF), platelet-derived growth factor (PDGF), or basic fibroblast growth factor (bFGF). Stromal cells expressed transcripts of both TIMP-1 and -2. Epithelial cells expressed TIMP-1, but little TIMP-2. TGF-beta did not regulate the expression of TIMP-1 or -2 in either stromal or epithelial cells. Our results suggest that the elevated levels of MMP-2 and -9 observed in prostate development and cancer may be due to the elevated TGF-beta associated with these tissues.
Wilson
Michael J
MJ
Minneapolis VA Medical Center, Minnesota 55417, USA. wilso042@tc.umn.edu
Sellers
Robert G
RG
Wiehr
Carol
C
Melamud
Ori
O
Pei
Duanqing
D
Peehl
Donna M
DM
D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000502:physiology; D006133:Growth Substances / Q000494:pharmacology; D006728:Hormones / Q000494:pharmacology; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D009361:Neoplasm Invasiveness / Q000503:physiopathology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D011467:Prostate / Q000166:cytology / Q000201:enzymology*; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000503:physiopathology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics*; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics*; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D006133:Growth Substances; D006728:Hormones; D019384:Nucleic Acid Synthesis Inhibitors; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D019715:Tissue Inhibitor of Metalloproteinase-1; D016212:Transforming Growth Factor beta; D019716:Tissue Inhibitor of Metalloproteinase-2; D003609:Dactinomycin; D003513:Cycloheximide; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9