Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jan 28, 2015; 21(4): 1125-1139
Published online Jan 28, 2015. doi: 10.3748/wjg.v21.i4.1125
Weichang’an and 5-fluorouracil suppresses colorectal cancer in a mouse model
Li Tao, Jin-Kun Yang, Ying Gu, Xin Zhou, Ai-Guang Zhao, Jian Zheng, Ying-Jie Zhu
Li Tao, Jin-Kun Yang, Ying Gu, Ai-Guang Zhao, Jian Zheng, Ying-Jie Zhu, Department of Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
Xin Zhou, Department of Pharmacy, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
Author contributions: Tao L and Yang JK designed the research; Tao L, Gu Y and Zhao AG performed the research; Zhu YJ and Zheng J analyzed the data; and Tao L, Yang JK and GuY, Zheng J wrote the paper; Zhou X performed preparation of the Chinese medicine and quality control.
Supported by Natural Science Research Fund of Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine; Budgetary Scientific Research Project of the Education Commission of Shanghai “The impact of Weichang’an decoction on β-catenin/MMP7 signaling pathway in nude mice with hepatic metastasis from colorectal cancer: a study on the molecular mechanism”, No. 2011JW33; Young Talent Scientific Research Project of Shanghai Municipal Commission of Health and Family Planning “Efficacy Evaluation of combined therapy with TCM and western medicine for the treatment of hepatic metastasis from unresectable colorectal cancer”, No. 20134y141.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Jin-Kun Yang, Chief physician, Professor, Department of Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, No. 725, South Wanping Road, Shanghai 200032, China. yangjinkun131229@163.com
Telephone: +86-21-64385700 Fax: +86-21-64398310
Received: May 6, 2014
Peer-review started: May 11, 2014
First decision: June 10, 2014
Revised: July 17, 2014
Accepted: August 13, 2014
Article in press: August 28, 2014
Published online: January 28, 2015
Abstract

AIM: To examine the effect of Weichang’an (WCA) and 5-fluorouracil (5-FU) on colorectal tumor and hepatic metastasis in a mouse model.

METHODS: Quantitative determination of hesperidin, the effective component in WCA decoction, was performed using HPLC. In vitro cytotoxicity of WCA was determined by treating HCT-116 cells with WCA diluents or serum extracted from rats that received WCA by oral gavage for 1 wk and MTT assays. Forty-eight nude mice received cecum implantation with tumor blocks subcutaneously amplified from human colon cancer cell line HCT-116. Mice were randomly divided into four treatment groups: control (CON), WCA, 5-FU and combination (WCA + 5-FU). Pathological examination of tumors consisted of tissue sectioning and hematoxylin and eosin staining. Tumor weight and size were measured, and the number of metastatic lesions was counted. Serum carcinoembryonic antigen (CEA) level was determined by ELISA. The expression levels of tumor genesis and metastasis-related proteins β-catenin and matrix metalloproteinase (MMP)-7 were measured by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR), immunohistochemistry and immunoblotting. Cell fractionation was used to investigate intracellular distribution of β-catenin.

RESULTS: Parenchymal tumors were palpable in the abdomens of nude mice 2 wk post-implantation and orthotopic tumor formation rate was 100% in all groups. 5-FU treatment alone significantly decreased tumor weight compared to the CON group (1.203 ± 0.284 g vs 1.804 ± 0.649 g, P < 0.01). WCA treatment alone reduced the rate of metastasis (50% vs 100%, P < 0.05). Combination treatment of WCA + 5-FU was the most effective, reducing the tumor weight (1.140 ± 0.464 g vs 1.804 ± 0.649 g, P < 0.01) and size (1493.438 ± 740.906 mm3vs 2180.259 ± 816.556 mm3, P < 0.05), the rate of metastases (40% vs 100%, P < 0.01), and serum CEA levels (31.263 ± 7.421 μg/L vs 43.040 ± 11.273 μg/L, P < 0.05). Expression of β-catenin and MMP-7 was decreased in drug-treated groups compared to controls, as detected using real-time quantitative RT-PCR, immunohistochemistry and immunoblotting, respectively. Cell fractionation assays revealed that nuclear translocation of β-catenin was reduced by WCA and/or 5-FU treatments.

CONCLUSION: Combination of WCA with 5-FU significantly inhibited colon tumor growth and hepatic metastases. Decreased expression of β-catenin and MMP-7 may be important.

Keywords: Colorectal cancer, Hepatic metastasis, Weichang’an formula, Orthotopic transplant nude mouse model, Chemotherapeutics 5-fluorouracil

Core tip: In this study, the anti-colon cancer activity of Weichang’an (WCA), a Chinese herbal medicine, was assessed in an orthotopic transplantation nude mouse model. Combination of WCA with 5-fluorouracil inhibited orthotopic tumor growth and hepatic metastases. Decreased expression of β-catenin and metalloproteinase-7, which are crucial proteins modulating tumor aggression, may be important for the anti-tumor properties of WCA.