Inhibitory effects of Chanling Gao on the proliferation and liver metastasis of transplanted colorectal cancer in nude mice.

Bing Yang,Feng-Xi Long,Jing-Yu Fan,Dong-Xin Tang,Jing-Yan Han,Zhu Yang,Quan Li,Chun-Shui Pan,Chuan-She Wang,Irina V Lebedeva

doi:10.1371/journal.pone.0201504

Abstract

This study aimed to explore the efficacy and mechanism of Chanling Gao (CLG), a compound Chinese medicine, on colorectal cancer (CRC). A model of transplanted CRC was established in nude mice. The mice were treated 7 days after CRC transplantation with either Capecitabine or CLG for 3 weeks. On the 28th day after the operation, CRC growth and liver metastasis were assessed by morphology, the changes in the expression of HIF-1α (hypoxia inducible factor-1α), stromal cell–derived factor-1 alpha (SDF-1α), CXCR4 (C-X-C chemokine receptor type 4), PI3K, and Akt in the transplanted tumor and SDF-1α and CXCR4 in the liver were detected by Western blot and immunohistochemistry. The protein contents of vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, and collagen IV in the serum and transplanted tumor and SDF-1α and CXCR4 in liver tissues were detected by enzyme-linked immunosorbent assay. In the Capecitabine and high dose CLG groups, the growth and liver metastasis of CRC were significantly inhibited, the protein levels of HIF-1α, SDF-1α, CXCR4, MMP-2, VEGF, PI3K, Akt, P-PI3K and P-Akt in the transplanted tumor were lower, while the content of collagen IV in the transplanted tumor was higher, than in Model group. A high dose of CLG inhibited the growth of transplanted tumor and liver metastasis of CRC in nude mice, probably by inhibiting the HIF-1α/SDF-1α-CXCR4/PI3K-Akt signaling pathway reducing the synthesis and release of VEGF and degradation of collagen IV.

Highlights

Colorectal cancer (CRC) is one of the most common malignant tumors, and its morbidity and mortality rank the third among all malignant tumors [1]
The F12K medium was purchased from Genview Scientific Inc. (Miami, FL, USA); PGL4.51 was purchased from Promega Corp. (Madison, WI, USA); Lipofectamine 2000 and Geneticin (G-418) were purchased from Life Technologies Corp. (Carlsbad, CA, USA); D-Luciferin was obtained from Gold Biotechnology (St Louis, MO, USA); Medical OB glue was purchased from Guangzhou Baiyun Chemical Industry Co., Ltd (Guangzhou, China); Chanling Gao (CLG) was provided by the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine (Guizhou, China); the Capecitabine tablets were purchased from Shanghai Roche Pharmaceuticals Ltd. (Shanghai, China); Antibodies against β-actin (1:2000; ab124964), CXCR4 (1:800; ab124824), SDF-1α (1:1000; ab25117), and HIF-1α(1:1000; ab16066) were purchased from Abcam Inc. (Cambridge, MA, USA)
CLG inhibits the growth of the transplanted tumor in CRC nude mice The results of luciferase activity assay showed that a stable cell line expressing luciferase (LoVo-luc cells) was established, and luminous intensity was proportional to cell density (Fig 1A)

Summary

Introduction

Colorectal cancer (CRC) is one of the most common malignant tumors, and its morbidity and mortality rank the third among all malignant tumors [1]. Distant metastasis is an important cause of poor prognosis in CRC patients, of which liver is the primary target organ for CRC hematogenous metastasis [2]. The liver metastases of most patients (80%–90%). Inhibitory effects of Chanling Gao on the proliferation and liver metastasis of transplanted CRC in nude mice analysis, decision to publish, or preparation of the manuscript

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Conclusion