Purchase this article with an account.
D.D. Klisovic, M.I. Klisovic, S.E. Katz, D. Effron, J. Wickham, M.R. Parthun, G. Marcucci; Histone Deacetylase Inhibitors (HDACIs), Inhibit Proliferation and Induce Apoptosis of Primary and Metastatic Uveal Melanoma Cell Lines in vitro: A Potential Therapeutic Strategy for Uveal Melanoma (UM) . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4977.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: UM is the most common primary malignant ocular tumor in adults. No effective chemotherapy regimens are available for either intraocular or metastatic UM. We tested the ability of HDACIs sodium butyrate (NaB), trichostatin (TSA) and depsipeptide (DP) to inhibit proliferation and induce apoptosis in UM cells in vitro. Methods: 3 primary and 2 metastatic (liver metastasis) UM cell lines were treated with NaB, TSA and DP for 48 hours. Cell proliferation was studied by MTT-based proliferation assay. Induction of apoptosis was studied by flow-cytometry. Changes in gene expression of Fas/FasL, p21 and p27 were studied by RT-PCR and changes in gene expression of Bcl-2/Bax by Real-time PCR. Histone acetylation (H3 and H4), Fas/FasL, p21, p27, bcl-2/bax and caspase-3 protein levels were determined by Western-blot and ELISA assayResults: Dose-dependent increase in histone (H3 and H4) acetylation was observed in all UM cell lines. Single pulse of HDACIs strongly inhibited cell growth in all primary and metastatic cell lines (range of 60-90% inhibition). Flow cytometry revealed significant induction of apoptosis (~ 3.8-fold increase). RT-PCR and Western-blot revealed dose-dependent up-regulation Fas/FasL, p21 and p27 proteins. A 5.6-fold increase in caspase-3 activity was observed. This increase could be inhibited by protein synthesis inhibitor cycloheximide. No changes in Bcl-2/Bax gene expression were detected by Real-time PCR or Western-blot. Conclusions: HDACIs are potent inhibitors of primary and metastatic UM cell growth in vitro. In addition, they strongly induce apoptosis in those cell lines. Our data suggest that inhibition of cell growth is mediated via the increase in p21 and p27 expression and induction of apoptosis via the Fas/FasL signaling pathway. Bcl-2/Bax signaling pathway does not seem to be involved in HDAC-induced apoptosis in UM cell lines. HDACIs may prove to be a valuable adjunctive treatment modality for primary and metastatic UM.
This PDF is available to Subscribers Only