Original Article
Copyright ©2011 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Aug 21, 2011; 17(31): 3605-3613
Published online Aug 21, 2011. doi: 10.3748/wjg.v17.i31.3605
Ginsenoside Rg3 inhibit hepatocellular carcinoma growth via intrinsic apoptotic pathway
Jian-Wen Jiang, Xin-Mei Chen, Xin-Hua Chen, Shu-Sen Zheng
Jian-Wen Jiang, Xin-Hua Chen, Shu-Sen Zheng, The Key Lab of Combined Multi-organ Transplantation, Ministry of Public Health, The Department of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China
Xin-Mei Chen, The Department of Pharmacy, Shandong Traditional Chinese Medicine University, Jinan 250355, Shandong Province, China
Author contributions: Jiang JW and Chen XM contributed equally to this work; Jiang JW, Chen XM, Chen XH and Zheng SS designed the research; Jiang JW, Chen XM and Chen XH performed the research and wrote the paper.
Supported by The National Natural Science Foundation of China, No. 30700778; the Health Bureau Fund of Zhejiang Province, No. 2007QN006, No. 2008B080 and No. 2008A050; and National Basic Research Program (973) of China, No. 2007CB513005
Correspondence to: Shu-Sen Zheng, PhD, MD, Department of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, School of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou 310003, Zhejiang Province, China. shusenzheng@zju.edu.cn
Telephone: +86-571-87236570 Fax: +86-571-87236466
Received: April 22, 2011
Revised: July 6, 2011
Accepted: July 13, 2011
Published online: August 21, 2011
Abstract

AIM: To investigate the anti-tumor function of ginsenoside Rg3 on hepatocellular carcinoma (HCC) in vitro and in vivo, and its mechanism.

METHODS: Hep1-6 and HepG2 cells were treated by Rg3 in different concentrations (0, 50, 100 and 200 μg/mL) in vitro. After incubation for 0, 6, 12, 24 and 48 h, cell viability was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Apoptosis was identified by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling. Caspase-3 activity was measured by chromophore p-nitroanilide and flow cytometry. Bcl-2 family proteins were ascertained by Western-blotting. Mitochondria membrane potential was detected by 5, 5’, 6’ 6’ - tetrachloro-1, 1’, 3, 3’ - tetraethylbenzimidazolylcarbocyanine iodide. Forty liver tumor-bearing C57Bl6 mice were divided randomly into 4 groups for intra-tumor injection of saline, ginsenoside Rg3, cyclophosphamide (CTX) and ginsenoside Rg3 + CTX combination.

RESULTS: The survival time was followed up to 102 d. The mice in the Rg3 + CTX group showed significant increased survival time compared with those in the control group (P < 0.05). Rg3 could inhibit HCC cell proliferation and induce cell apoptosis in vitro in the concentration and time dependent manner. It also induced mitochondria membrane potential to decrease. Caspase-3 activation can be blocked by the inhibitor z-DEVD-FMK. Bax was up-regulated while Bcl-2 and Bcl-XL were down-regulated after Rg3 treatment.

CONCLUSION: Our data suggested that Rg3 alone or combined with CTX inhibited tumor growth in vivo and prolonged mouse survival time by inducing HCC cell apoptosis via intrinsic pathway by expression alterations of Bcl-2 family proteins.

Keywords: Ginsenoside Rg3, Apoptosis, Hepatocellular Carcinoma, Bcl-2 family proteins, Cyclophosphamide