Romidepsin reduces histone deacetylase activity, induces acetylation of histones, inhibits proliferation, and activates apoptosis in immortalized epithelial.

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Presentation transcript:

Romidepsin reduces histone deacetylase activity, induces acetylation of histones, inhibits proliferation, and activates apoptosis in immortalized epithelial endometriotic cells Patrick Imesch, M.D., Daniel Fink, M.D., André Fedier, Ph.D. Fertility and Sterility Volume 94, Issue 7, Pages 2838-2842 (December 2010) DOI: 10.1016/j.fertnstert.2010.04.052 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Expression of class I (HDAC-1, HDAC-2, HDAC-3, HDAC-6) and class II (HDAC-4, HDAC-5) histone deacetylase (HDAC) isoforms in 11z immortalized epithelial endometriotic cells (A) and effects of romidepsin (Romi) on the HDAC enzymatic activity (B), histone acetylation (C), p21 and p27 expression (D), cyclin B1 and cyclin D1 expression (E), proliferation (F), and apoptosis activation expressed as the cleavage of the full-length 47-kD procaspase-9 into its 37-kD fragment, the full-length 32-kD procaspase-3 into its 17-kD fragment (G), and the full-length 116-kD poly (ADP-ribose) polymerase (PARP) 1 precursor into its 89-kD fragment (H). (A, D, E) Protein expression, (C) accumulation of acetylated histones, and (G, H) proteolytic cleavage of caspases and PARP-1 precursors were determined by immunoblot analysis of lysates from 11z cells treated with romidepsin (concentrations and treatment duration as indicated). β-Actin is the sample loading control. (D) A cell lysate of HCT116 colon tumor cells treated with 15 μmol/L SAHA was used as positive control for p27 (posCo). Data are representatives of two independent data sets. (B) Romidepsin-induced reduction of HDAC enzymatic activity in 11z cells was determined colorimetrically using an HDAC activity assay kit and is presented as the relative values of the enzymatic activities (% of untreated control). HDAC enzymatic activity refers to the relative values of measured optical density per milligram of protein. Also shown are the positive (PosCo; HeLa cell nuclear extract) and the negative (NegCo; HeLa cell nuclear extract plus 20 μmol/L trichostatin A [TSA]) assay controls. Data points are the mean ± SD of two independent experiments. (F) Proliferation inhibition of 11z cells treated with romidepsin (concentrations and treatment duration as indicated) was assessed by the methylthiazolyl tetrazolium assay. Data (mean ± SD of three independent experiments) are presented as relative proliferation as a function of romidepsin concentration and treatment duration. ∗P<.05 for the respective treated cultures vs. control (two-tailed Student t test; P<.05 was considered to be statistically significant). Fertility and Sterility 2010 94, 2838-2842DOI: (10.1016/j.fertnstert.2010.04.052) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions