Abstract
Abstract: :
Purpose. To study the expression of MMP–2 and MMP–9 of cultured human uveal melanocytes, and to test the effects of basic fibroblast growth factor (bFGF) and 12–0–tetradecanoyl–phorbol–13–acetate (TPA) on the expression of these MMPs. Methods: Human uveal melanocytes were isolated and cultured from donor eyes as previously reported (Invest Ophthalmol Vis Sci 33:2210, 1993). Early passages of 4 cell lines of uveal melanocytes were plated into multi–well plates and cultured with or without bFGF (20 ng/ml) and TPA (10 ng/ml). After 48 hours, the conditioned culture medium was collected for gelatin zymography. The cells were detached, the mRNA was extracted and the amount of MMP mRNA was analyzed by the RT–PCR using two primers specific for human MMP–2 and MMP–9. Results:In cells cultured with the standard culture medium, gelatin zymography of conditioned culture medium detected MMP–2 (72 kDa) and a relatively small amount of MMP–9 (92 kDa), both in the latent form. RT–PCR analysis also revealed the MMP–2 and MMP–9 mRNA in cultured uveal melanocytes. Addition of TPA to the culture medium stimulated the production of MMP–2 (170% of the control) and MMP–9 (626% of the control) and the transcription of MMP–2 (139% of the control) and MMP–9 (217% of the control) by uveal melanocytes (P < 0.01), whereas the addition of bFGF did not showed significant effects of the expression of these MMPs (P > 0.05). Conclusions: Human uveal melanocytes express the MMP–2 and MMP–9 in vitro. TPA, but not bFGF, stimulates the expression of MMP–2 and MMP–9 by uveal melanocytes.
Keywords: melanocytes • uvea • enzymes/enzyme inhibitors