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Rinat Dror, Michal Lederman, Kazuo Umezawa, Vivian Barak, Jacob Pe'er, Itay Chowers; Characterizing the Involvement of the Nuclear Factor-kappa B (NFκB) Transcription Factor in Uveal Melanoma. Invest. Ophthalmol. Vis. Sci. 2010;51(4):1811-1816. doi: https://doi.org/10.1167/iovs.09-3392.
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© ARVO (1962-2015); The Authors (2016-present)
To examine the involvement of nuclear factor-kappa B (NFκB) pathways in uveal melanoma (UM) and to assess their potential as a therapeutic target for metastatic UM.
Samples from primary (n = 7) and metastatic (n = 7) UM were evaluated for NFκB transcription factor family expression by quantitative PCR (QPCR), immunofluorescent staining, and Western blot analysis. The effect of two NFκB inhibitors, DHMEQ and BMS-345541, on two cell lines derived from UM liver metastases was assessed. Cell proliferation was examined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, methylene blue assay, and immunostaining for Ki-67. Apoptosis was assessed by immunostaining for activated caspase 3.
NFκB1, NFκB2, RelA, RelB, and NIK were expressed in primary UM and in its liver metastases. NFκB2, RelB, and NIK showed significantly higher mRNA levels in metastases from UM compared with primary tumors (3.4-fold, P = 0.03; 3.6-fold, P = 0.05; 3.5-fold, P = 0.03; respectively). NFκB2 protein activation was 3.9-fold higher in metastases (P = 0.03). NFκB inhibition reduced metastatic cell proliferation by 9.2-fold and 1.9-fold according to Ki67 staining (P = 0.04) and methylene blue assay (P = 6 × 10−7), respectively. Both NFκB inhibitors achieved dose-dependent reductions of UM cell proliferation in both cell lines (P < 0.001). NFκB inhibition resulted in a 6.3-fold increase of apoptosis (P = 7 × 10−7).
These data indicate that the NFκB1 and NFκB2 pathways are active in both primary and metastatic UM and that these pathways regulate metastatic cell proliferation and apoptosis. The role of NFκB as a therapeutic target for UM should be further evaluated.
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