GTPs and SFN altered epigenetic enzymes expression and their activity in ERα-negative breast cancer cells.

Breast cancer MDA-MB-231 cells were treated with the indicated concentration of GTPs and SFN alone or in combinations of both for 3 days. Nuclear extracts were prepared and 20 µg of protein was used to estimate DNMTs (panel A), HDAC and HATs (panel B) activities using the colorimetric assay kit as described under Materials and Methods. Non-treated MCF-7 cells were used as a positive control. Values are representative of three independent experiments and represented as percent control ± SD; statistical significance against nontreated MDA-MB-231 control, ^*P<0.05. C) Effect of GTPs and SFN alone or in combinations of both on DNMTs, SUV39H1 and HDACs expression in ERα-negative human breast cancer MDA-MB-231 cells. Cell lysates were prepared at 3 days of post-treatments at the indicated doses followed by western blotting to analyze DNMTs (DNMT1, DNMT3a and DNMT3b), HDACs (HDAC1, HDAC4 and HDAC6) and SUV39H1 expression. Non-treated MCF-7 cells were used as a positive control. Actin was used as an equal loading control. D) Graphical representations are indicative of relative band intensity of DNMTs expression in MDA-MB-231 cells, normalized with β-actin. Values are mean band intensity of three independent blot ± SD.