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Shahar Frenkel, Jacob Pe'er; A New in vivo Model for Testing Treatments for Liver Metastases of Uveal Melanoma: Model Construction and Use for Testing NFkB Inhibition. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3422.
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© ARVO (1962-2015); The Authors (2016-present)
To describe a new animal model for in vivo testingof treatments for liver metastases of uveal melanoma, and its use for testing an NFkB inhibitor.
A metastatic uveal melanoma cell line (C918) was transfected with the luciferase gene in a stable transfection. The new C918-Luc cell line was trypsinized, washed in 1xPBS and injected directly into the livers of C57Bl mice (for the model construction) or SCID mice (for testing the NFkB inhibitor) through a small (1cm) abdominal wall incision via a 0.5cc insulin syringe (29 gauge needle) in a non-reflux technique. Cells were allowed to settle for 1 week before administration of the BMS-345541 NFkB inhibitor. Intra-hepatic cells were visualized with an IVIS bioluminescence camera (Caliper Life Sciences, Hopkinton, MA USA) 10 minutes after injecting the mice with luciferin (IP 300mg/0.3cc). BMS-345541 (5mg/kg) was administered by oral gavage 3 times a week for 3 weeks, followed by euthanasia and harvesting of the livers for histopathologic evaluation. Bioluminescence was measured twice a week. The research reported here was conducted in compliance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.
Bioluminescence was detectable in the liver immediately post op, increased over a period of one week in C57Bl mice and then decreased and disappeared over another week. In SCID mice, bioluminescence started decreasing after administration of the inhibitor. Experiments are not finished yet. Final results of the in vivo effect of NFkB inhibition will be presented at the meeting.
Combining the direct injection model with bioluminescence provides a way to follow the effect of anti-metastatic treatments in vivo. Inhibition of NFkB shows potential as a new treatment for metastatic uveal melanoma.
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