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A novel tripeptide, tyroserleutide, inhibits irradiation-induced invasiveness and metastasis of hepatocellular carcinoma in nude mice

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Summary

Previous studies have demonstrated that tyroserleutide (YSL) inhibits tumor growth in an animal model of hepatocellular carcinoma (HCC). However, its effects on HCC metastasis are still not fully understood. To examine YSL as a novel agent to prevent HCC metastasis, a metastatic human HCC orthotopic nude mouse model of MHCC97L was used. The antitumor and antimetastasis effects of YSL were also evaluated in combination with radiation. Hypoxia and epithelial-mesenchymal transition (EMT)-related molecules were studied. YSL inhibited MHCC97L cell invasion in vitro with or without irradiation. YSL did not significantly inhibit tumor growth but decreased pulmonary metastasis and prolonged life-span for more than 40 days, which correlated with down-regulation of matrix metalloproteinase-2. Radiotherapy inhibited early-stage tumor growth and promoted tumor hypoxia. The re-implanted tumor volume in the radiotherapy group was not significantly different from the control, in which the incidence of lung metastasis increased after radiotherapy (6/6 versus 3/6, P = 0.046); however, YSL inhibited the growth of re-implanted tumor after radiotherapy. Furthermore, YSL at 160 or 320 μg/kg/day almost completely inhibited lung metastasis induced by irradiation (1/6 versus 6/6, P = 0.002 for both dosages). YSL down-regulated hypoxia-inducible factor 1α (HIF-1α) and transmembrane protease serine 4 (TMPRSS4), and inhibited EMT was associated with the antimetastasis capability of YSL. Our data suggest that YSL inhibits the enhanced invasiveness and metastatic potential of HCC induced by irradiation through down-regulation of HIF-1α and TMPRSS4 and inhibition of EMT. YSL may have potential as a new antimetastasis agent for radiotherapy.

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Abbreviations

EMT:

epithelial-mesenchymal transition

HCC:

hepatocellular carcinoma

HIF-1α:

hypoxia-inducible factor 1α

MMP-2:

matrix metalloproteinase-2

TMPRSS4:

transmembrane protease serine 4

YSL:

tyroserleutide

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Acknowledgements

The authors thank Dr. Zhen-Yu Zhang (Radiation Oncology, Zhongshan Hospital, Fudan University, Shanghai, China) for offering radiotherapy helps. This study was jointly supported by a China Postdoctoral Science Foundation–funded project (No. 20080440077 and 200902203) and a National Key Science and Technology Specific project (2008ZX10002-021).

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Authors and Affiliations

  1. Liver Cancer Institute and Zhongshan Hospital, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University, 136 Yi Xue Yuan Road, Shanghai, 200032, China

    Jin-Bin Jia, Wen-Quan Wang, Hui-Chuan Sun, Liang Liu, Xiao-Dong Zhu, Ling-Qun Kong, Zong-Tao Chai, Wei Zhang, Ju-Bo Zhang, Hua-Xiang Xu, Wei-Zhong Wu, Lu Wang & Zhao-You Tang

  2. Shenzhen Kangzhe Pharmaceutical Co. Ltd., Shenzhen, 518057, China

    Jin-Bin Jia

  3. Radiation Oncology, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Zhao-Chong Zeng

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  1. Jin-Bin Jia
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Correspondence to Zhao-You Tang.

Additional information

Jin-Bin Jia and Wen-Quan Wang contributed equally to this work.

Jin-Bin Jia works for Shenzhen Kangzhe Pharmaceutical Co. Ltd. which owns the patent for tyroserleutide. The other authors have no financial disclosures or conflicts of interest to report.

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Fig. S1

Reference markers of tumor hypoxia pimonidazole staining and HIF-1α expression in tumor tissue with or without irradiation treatment. Low (a) or high (b) pimonidazole staining, (50×). HIF-1α mRNA (c) and protein (d) level in control and radiotherapy group. *P < 0.05, compared with the control. β-actin was an internal control in Western blot analysis. (GIF 63 kb)

High resolution image (TIFF 4489 kb)

Fig. S2

Effects of YSL on weight loss in nude mice after radiotherapy. (GIF 32 kb)

High resolution image (TIFF 39 kb)

Fig. S3

Effects of YSL on the expression of HIF-1α and TMPRSS4 analyzed by immunohistochemistry. a–e HIF-1α staining was mainly on the nuclei or cytoplasm of tumor cells. f–j TMPRSS4 staining was mainly on the cytomembrane of tumor cells. a and f showed high expression of the two molecules, and the others showed low expression of the two molecules. (GIF 613 kb)

High resolution image (TIFF 4803 kb)

Table S1

The differential expression of four tumor invasion and metastasis-related genes in tumor tissue different times after radiotherapy. (DOC 35 kb)

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Jia, JB., Wang, WQ., Sun, HC. et al. A novel tripeptide, tyroserleutide, inhibits irradiation-induced invasiveness and metastasis of hepatocellular carcinoma in nude mice. Invest New Drugs 29, 861–872 (2011). https://doi.org/10.1007/s10637-010-9435-1

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