TY - JOUR
T1 - Suppression of Vascular Endothelial Growth Factor via Inactivation of Eukaryotic Elongation Factor 2 by Alkaloids in Coptidis rhizome in Hepatocellular Carcinoma
AU - Tan, Hor Yue
AU - Wang, Ning
AU - Tsao, Sai-Wah
AU - Zhang, Zhangjin
AU - Feng, Yibin
N1 - Funding Information:
The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported in part by grants from the research council of the University of Hong Kong (project code 104002320), Research Grant Committee of Hong Kong SAR of China (General Research Fund, project code 10500362), and Wong’s donation for modern oncology of Chinese medicine (project code 20006276).
Publisher copyright:
© The Author(s) 2013
PY - 2014/9
Y1 - 2014/9
N2 - Aim of study: To investigate the inhibitory effect of Coptidis rhizome aqueous extract (CRAE) on vascular endothelial growth factor (VEGF) expression and tumor angiogenesis in hepatocellular carcinoma (HCC). Methods: Quality control of CRAE was determined. Secretion of VEGF protein and expression of its mRNA in MHCC97L and Hep G2 cells were measured with enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction. Synthesis of nascent protein was determined by AHA-protein-labeling technologies. The in vivo antiangiogenic effect of CRAE was evaluated with a xenograft model. Results: Absence of organochlorine pesticides in CRAE was found, and phytochemical analysis showed that its components were in proportion of magnoflorine 2.2%, jatrorrhizine 1.68%, palmatine 4.4%, and berberine 13.8%. CRAE exhibited significant inhibition on VEGF secretion from MHCC97L and HepG2 cells at nontoxic doses. The mRNA transcripts of VEGF could not be inhibited by CRAE; however, synthesis of VEGF nascent protein was potently blocked by CRAE. CRAE intervention increased the phosphorylation of eukaryotic elongation factor 2 (eEF2) in HCC cells, which blocked eEF2 activity for proceeding nascent protein synthesis. The activity of eEF2 was restored in CRAE-treated HCC cells in the presence of A484594, leading to the recovery of VEGF expression. Berberine was found to be the major active component in CRAE; however, CRAE is more effective in inhibiting eEF2 activity compared to berberine treatment alone, suggesting the additive effect of other components present. Reduction of tumor size and neovascularization were observed in mice xenograft model. Conclusion: Our study postulates the antiangiogenic effect of CRAE on hepatocellular carcinoma via an eEF2-driven pathway.
AB - Aim of study: To investigate the inhibitory effect of Coptidis rhizome aqueous extract (CRAE) on vascular endothelial growth factor (VEGF) expression and tumor angiogenesis in hepatocellular carcinoma (HCC). Methods: Quality control of CRAE was determined. Secretion of VEGF protein and expression of its mRNA in MHCC97L and Hep G2 cells were measured with enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction. Synthesis of nascent protein was determined by AHA-protein-labeling technologies. The in vivo antiangiogenic effect of CRAE was evaluated with a xenograft model. Results: Absence of organochlorine pesticides in CRAE was found, and phytochemical analysis showed that its components were in proportion of magnoflorine 2.2%, jatrorrhizine 1.68%, palmatine 4.4%, and berberine 13.8%. CRAE exhibited significant inhibition on VEGF secretion from MHCC97L and HepG2 cells at nontoxic doses. The mRNA transcripts of VEGF could not be inhibited by CRAE; however, synthesis of VEGF nascent protein was potently blocked by CRAE. CRAE intervention increased the phosphorylation of eukaryotic elongation factor 2 (eEF2) in HCC cells, which blocked eEF2 activity for proceeding nascent protein synthesis. The activity of eEF2 was restored in CRAE-treated HCC cells in the presence of A484594, leading to the recovery of VEGF expression. Berberine was found to be the major active component in CRAE; however, CRAE is more effective in inhibiting eEF2 activity compared to berberine treatment alone, suggesting the additive effect of other components present. Reduction of tumor size and neovascularization were observed in mice xenograft model. Conclusion: Our study postulates the antiangiogenic effect of CRAE on hepatocellular carcinoma via an eEF2-driven pathway.
KW - hepatocellular carcinoma
KW - Coptidis rhizome aqueous extract
KW - vascular endothelial growth factor
KW - eukaryotic elongation factor 2
KW - nascent protein synthesis
UR - http://www.scopus.com/inward/record.url?scp=84906053371&partnerID=8YFLogxK
U2 - 10.1177/1534735413513635
DO - 10.1177/1534735413513635
M3 - Journal article
C2 - 24363282
AN - SCOPUS:84906053371
SN - 1534-7354
VL - 13
SP - 425
EP - 434
JO - Integrative Cancer Therapies
JF - Integrative Cancer Therapies
IS - 5
ER -
