Original Article
The inhibitory effect of Withaferin A on the growth of orthotopic xenograft tumor of hepatocellular carcinoma in nude mice and the mechanism
Mu Xianmin, Shi Wei, Xu Yue, Hu Shi, Yang Jing, Xu Che, Zhang Chen, Pan Jinshun, Geng Biao, You Qiang
Published 2017-12-20
Cite as J Chin Physician, 2017,19(12): 1800-1803,1806. DOI: 10.3760/cma.j.issn.1008-1372.2017.12.012
Abstract
ObjectiveTo investigate the inhibitory effect of Withaferin A (WFA) on the growth of orthotopic xenograft tumor of hepatocellular carcinoma in nude mice and the mechanism of its antitumoral effect.
MethodsFor in vivo model, anti-tumor efficacy of Withaferin A was evaluated in nude mice models of human liver cancer orthotopic xenograft. The nude mice were randomly divided into model group, Sunitinib group, and Withaferin A groups [6, 3 mg/(kg·d)]. All mice were given intraperitoneal injection for 14 days. Tumor volume and tumor weight were observed. Antiangiogenic effects were assessed in vivo by the tumor inhibition rate and microvessel density. Quantitative polymerase chain reaction (QPCR) assay was used to detect the mRNA expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiopoietin-2 (Ang-2), vascular endothelial growth factor receptor 2 (VEGFR2) from tumor tissues. For in vitro experiments, the cell count kit 8 (CCK8) assay was used to detect the effect of Withaferin A on HepG2 cells proliferation. QPCR assay and enzyme-linked immunosorbent assay (ELISA) were used to detect the mRNA expression of VEGF.
ResultsCompared to the model group, the high-dose Withaferin A group and the Sunitinib group had a significantly lower tumor weight (P<0.05). The tumor inhibition rate was 42.69% in the high-dose Withaferin A group, 20.22% in the low-dose Withaferin A group, and 49.43% in the Sunitinib group. The growth of HepG2 cells was significantly inhibited by different concentrations of Withaferin A, and the 50% concentration of inhibition (IC50) of Withaferin A were (2.64±0.18)μmol/L at 24 h. Withaferin A (6, 3 μmol/L) could inhibit the protein and mRNA expression of VEGF (P<0.05).
ConclusionsWithaferin A significantly reduces the growth of orthotopic xenograft tumor of hepatocellular carcinoma in nude mice via antiangiogenic effect. Downregulation of the protein and mRNA expression of VEGF by WFA may be one mechanism of its anti-liver cancer effect.
Key words:
Withanolides/PD; Liver neoplasms/ZD; Cell proliferation/DE; Neovascularization, pathologic
Contributor Information
Mu Xianmin
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
Shi Wei
Department of Drug Screening and Evaluation, Chia Tai Tianqing Pharmaceutical Group Co., Ltd, Nanjing 210023, China
Xu Yue
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
Hu Shi
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
Yang Jing
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
Xu Che
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
Zhang Chen
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
Pan Jinshun
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
Geng Biao
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
You Qiang
Department of Biotherapy, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
