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Analysis of Angiostatin Activity and Angiostatin Treatment in a Murine-model for Metastatic Ocular Melanoma
1Dithmar S., 2Yang H., 3Grossniklaus H. E.,
1Ruprecht-Karls-Universität Heidelberg, Universitäts-Augenklinik (Heidelberg)
2Emory University, Department of Ophthalmology (Atlanta)
3Emory University School of Medicine, Department of Ophthalmology (Atlanta)
Purpose: To analyse the angiostatin activity of B16-LS9 melanoma cells and to perform an experimental trial for the effect of angiostatin (given after enucleation) on metastases and micrometastases in a murine uveal melanoma model.
Method: The angiostatin assay was performed as previously described (Apte et al, Arch Ophthalmol 2001;119:1805). B16-LS9 tissue culture melanoma cells were inoculated into the posterior compartment of two groups of C57BL6 mice. The inoculated eyes were enucleated at 7 days and the mice were sacrificed at 21 days post-inoculation with number of metastases and micrometastases determined from the necropsy using standard techniques. Group 1 (n=22) was the control group, group 2 (n=24) was the treatment group. The groups were given daily SC injections of 50ml of PBS (group 1) or human recombinant angiostatin (0.6mg/ml)(group 2) for 14 days starting day 1 post-enucleation.
Results: B16-LS9 melanoma cells produce angiostatin (angiostatin activity: 1.27%). Pulmonary metastases were detected in 3/22 (14%) and 2/24 (12,5%) of groups 1 and 2, respectively. Hepatic micrometastases appeared in 12/22 (55%) and 8/24 (33%) of groups 1 and 2, respectively.
Conclusions: Experimental studies have shown that recombinant angiostatin inhibits angiogenesis and metastatic cancer. The removal of an angiostatin producing primary tumor (enucleation) might support the development of metastases due to the lack of angiostatin production. This experimental study supports the hypothesis that angiostatin treatment can decrease the metastatic rate.
Supported by NIH EY13165, NIH EY03060 and RPB Inc.