Aberrant Regulation of HDAC2 Mediates Proliferation of Hepatocellular Carcinoma Cells by Deregulating Expression of G1/S Cell Cycle Proteins
Figure 6
Sustained suppression of HDAC2 attenuates the tumorigenic potential of Hep3B cells in vitro and in vivo.
(A) Confirmation of HDAC2 suppression by its specific regulation of cell cycle components in HDAC2 depleted cell lines. A typical result of three performed experiments is shown. (B) Time course cell counting analyses of Hep3B and two stable cell lines (Hep3B_Mock and Hep3B_HDAC2KD). The cell counts show the results of four independent experiments. (C) HDAC2-deficient (Hep3B_HDAC2KD) or Mock (Hep3B_Mock) stable cell line was blocked in G2/M transition by nocodazole and then released in fresh medium. The DNA content was determined by FACS analysis with PI-stained cells at each time point (0, 2, 6, 12, 24, 48 h after release) (left). The percentage of cells in G1 phase was calculated and represented as bar graph (right). A typical result of three performed experiments is shown. (D) In vitro Colony formation assay showed the effect of HDAC2 inhibition on the growth of cell colonies after three weeks incubation. Quantification of colony numbers shown is mean ± SD of three independent experiments (top). The representative scan of 0.5% crystal violet-stained cells (bottom). (E) Tumor growth of Hep3B cell xenografts. (F) Representative tumors obtained at sacrifice after 52 days of growth (upper left) and mice (upper right). The tumor volume at sacrifice was presented as bar graph. Values shown are mean ± SD, n = 5 for Mock; n = 3 for HDAC2KD. Data in (B, D, E and F) were analyzed by Student's t-test. * p<0.05, Hep3B_HDAC2KD vs Hep3B_Mock.