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C.M. Cebulla, M.–E. Jockovich, L. Bajenaru, T.G. Murray; Gelatinase Activity in Human Retinoblastoma and Choroidal Melanoma . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2805.
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We have previously demonstrated that the gelatinases matrix metalloproteinase (MMP)–2 and –9 are upregulated in the LHßTag transgenic mouse model of retinoblastoma. We tested the hypothesis that gelatinase activity is similarly upregulated in human retinoblastoma and choroidal melanoma tumors.
To evaluate gelatinase activity, in situ zymography was performed. Portions of enucleation specimens were removed after surgery and immediately snap frozen and sectioned into 5 micron sections. Eight eyes with retinoblastoma and three with malignant choroidal melanoma were obtained. A negative control eye without tumor was obtained from the Florida Lions Eye Bank. The sections were evaluated using the Molecular Probes EnzChek fluorescent gelatinase assay. Sections were exposed to DQ gelatin for 13–18 hours or buffer as a negative control. Sections were rinsed with PBS, counter–stained with DAPI and coverslipped with antifade mounting reagent. The fluorescence resulting from gelatinase cleavage of DQ gelatin was evaluated with an Olympus fluorescence microscope.
In situ zymography demonstrates that gelatinase activity is upregulated in both retinoblastoma and choroidal melanoma tumors compared to negative controls. The gelatinase activity was highest along vascular–like structures within the tumor and along the edge of the tumor in both retinoblastoma and melanoma.
Similar to animal studies, gelatinase activity is upregulated in human retinoblastoma and choroidal melanoma tumors. Gelatinase activity is most intense in vascular structures, demonstrating its importance in angiogenesis and as a potential target in adjuvant therapy design.
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