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Evidence-Based Complementary and Alternative Medicine
Volume 2013, Article ID 185158, 11 pages
http://dx.doi.org/10.1155/2013/185158
Research Article

Synergistic Anticancer Effects of Vorinostat and Epigallocatechin-3-Gallate against HuCC-T1 Human Cholangiocarcinoma Cells

National Research and Development Center for Hepatobiliary Disease, Pusan National University Yangsan Hospital, Gyeongnam 626-770, Republic of Korea

Received 8 January 2013; Accepted 28 May 2013

Academic Editor: Andreas Sandner-Kiesling

Copyright © 2013 Tae Won Kwak et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Figure 1 showed that the synergistic effect of vorinostat/EGCG combination on the proliferation and viability of HuCC–T1 cells. Combination of vorinostat and EGCG showed synergistic growth inhibition and cytotoxic effect against HuCC–T1 cells. Furthermore, less than 10 % of cells were remained 72 h after treatment of vorinostat/EGCG combination.

Supplementary Figure 2 showed the cell cycle arrest analysis of HuCC–T1 cells after treatment of vorinostat/EGCG combination. As shown in supplementary Figure 2, G1 phase was decreased and sub–G1 phase increased after treatment of vorinostat/EGCG combination.

Supplementary Figure 3 showed the p53 expression of HuCC–T1 cells after treatment of vorinostat/EGCG combination. Compared to single treatment of vorinostat or EGCG, combination of vorinostat/EGCG clearly enhanced p53 nuclear translocation in Hucc–T1 cells, indicating that combination of vorinostat/EGCG induced apoptosis of HuCC–T1 cells.

Supplementary Figure 4 showed the MMP2 and 9 expression of HuCC–T1 cells after treatment of vorinostat or EGCG. Both vorinostat and EGCG dose‐dependently suppressed MMP–2 expression of HuCC–T1 cells even though vorinostat did not significantly affected MMP–9 expression of HuCC–T1 cells.

Supplementary Figure 5 showed the ROS generation after treatment of vorinostat and EGCG. As shown in supplementary Figure 5, vorinostat dose–dependently increased ROS generation of HuCC–T1 cells. However, EGCG significantly decreased ROS generation of HuCC–T1 cells until 200 min and then the level of ROS was slightly changed.

Interestingly, the ROS production of HuCC–T1 cells at treatment of vorinostat/EGCG combination was quite similar to the results of EGCG, indicating that EGCG has dominant effect on the ROS generation in the HuCC–T1 cells compared to vorinostat. These results indicated that vorinostat and EGCG combination showed synergistic effect on the proliferation, viability, cell cycle, apoptosis, MMP expression, and ROS generation of HuCC–T1 cells.

  1. Supplementary Material