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ASPP2 suppresses stem cell-like characteristics and chemoresistance by inhibiting the Src/FAK/Snail axis in hepatocellular carcinoma

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Tumor Biology

Abstract

Hepatocellular carcinoma (HCC) is the third leading cause of death in cancer patients worldwide. Understanding the molecular pathogenesis of HCC recurrence and chemoresistance is key to improving patients’ prognosis. In this study, we report that downregulation of ASPP2, a member of the ankyrin-repeat-containing, SH3-domain-containing, and proline-rich-region-containing protein (ASPP) family, bestowed HCC cells with stem-like properties and resistance to chemotherapy, including the expansion of side population fractions, formation of hepatospheroids, expression of stem cell-associated genes, loss of chemosensitivity, and increased tumorigenicity in immunodeficient mice. An expression profiling assay revealed that ASPP2 specifically repressed focal adhesion kinase (FAK)/Src/extracellular signal regulated kinase (ERK) signaling. ASPP2 does this by physically interacting with C-terminal Src kinase (CSK) and stimulating its kinase activity, which eventually leads to activator protein 1 (AP1)-mediated downregulation of Snail expression. In addition, pharmacologic inhibition of Src attenuated the effects of ASPP2 deficiency. Our findings present functional and mechanistic insight into the critical role of ASPP2 in the inhibition of HCC stemness and drug resistance and may provide a new strategy for therapeutic combinations to treat HCC.

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Abbreviations

EMT:

Epithelial-mesenchymal transition

GSEA:

Gene set enrichment analyses

IF:

Immunofluorescence

IP:

Immunoprecipitation

KEGG:

Kyoto Encyclopedia of Genes and Genomes

qRT-PCR:

Quantitative reverse-transcription polymerase chain reaction

shRNA:

Short hairpin RNA

SP:

Side population

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Acknowledgments

This work is supported in part by grants from Ministry of Science and Technology of China “973”and “863”programs (2014AA021103, 2014AA020704, 2010CB945600), National Nature Science Foundation of China (81472719, 81301781, 81402539), Beijing Municipal Commission of Science and Technology, Shanghai Key Laboratory of Cell Engineering (14DZ2272300), and Shanghai Leading Academic Discipline Project (B905).

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    Authors and Affiliations

    1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, China

      Lu Xu

    2. PLA General Hospital Cancer Center Key Lab, PLA Postgraduate School of Medicine, 28 Fuxing Road, Beijing, China

      Xin Tong, Sujie Zhang, Fan Yin, Xiaoyan Li, Huafeng Wei, Cheng Li & Jian Zhao

    3. Beijing Key Laboratory of Cell Engineering and Antibody, Beijing, China

      Xin Tong, Sujie Zhang, Fan Yin, Xiaoyan Li, Huafeng Wei, Cheng Li & Jian Zhao

    4. State Key Laboratory of Antibody Medicine and Targeting Therapy, Shanghai, China

      Yajun Guo

    5. International Joint Cancer Institute, The Second Military Medical University, 800 Xiang Yin Road, New Building 9th Floor, Shanghai, China

      Jian Zhao

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    1. Lu Xu
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    Correspondence to Jian Zhao.

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    The authors declare that all procedures performed in this study involving animals were approved by Institutional Animal Care and Use Committee (IACUC) of Second Military Medical University and PLA General Hospital.

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    Lu Xu and Xin Tong contributed equally to this work.

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    Xu, L., Tong, X., Zhang, S. et al. ASPP2 suppresses stem cell-like characteristics and chemoresistance by inhibiting the Src/FAK/Snail axis in hepatocellular carcinoma. Tumor Biol. 37, 13669–13677 (2016). https://doi.org/10.1007/s13277-016-5246-0

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