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. 2013 Nov 1;8(11):e78417. doi: 10.1371/journal.pone.0078417

Figure 3. Restoration of expression loss of RhoB gene.

Figure 3

Regulation of RhoB expression loss by histone deacetylase (HDAC) inhibitor trichostatin (TSA) and methyltransferase inhibitor 5-Azacytidine (5-Aza). (A) Semi-quantitative RT-PCR analysis of RhoB expression levels in ovary carcinoma cell: Cells were either nutreated (NT), treated with 5-Aza at 2 μM, with TSA at 0.5 μM, or with combination of TSA (0.5 μM) and 5-Aza (2 μM). It can be seen that 5-Aza did not increase RhoB expression. However, TSA could significantly reverse RhoB silencing in ovary cancer SKOV3 and A2780 cells. β-Actin was used as inner control. (B) The expression confirmation of RhoB by Western blotting analysis: SKOV3 and A2780 cells were exposed to 0.5μM TSA at different time points. β-actin was used as equal loading control. As shown, TSA treatment, but not 5-Aza, revived the protein expression of RhoB. (C) TSA-mediated cell growth arrests of ovary cancer cells: SKOV3 and A2780 cells were treated with gradient TSA for 10, 24 and 48 hours. The inhibition rate was calculated as a percentage of the in vitro proliferation in terms of untreated control cells by MTT. TSA inhibits SKOV3 and A2780 cells growth in a concentration- and time-dependent manner. The results were expressed as mean±S.D.