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Pyruvate kinase M2 promotes pancreatic ductal adenocarcinoma invasion and metastasis through phosphorylation and stabilization of PAK2 protein

Oncogenevolume 37pages17301742 (2018) | Download Citation

Abstract

Pyruvate kinase muscle isozymes (PKMs) have crucial roles in regulating metabolic changes during carcinogenesis. A switch from PKM1 to PKM2 isoform was thought to lead to aerobic glycolysis promoting carcinogenesis, and was considered as one of the cancer signatures. However, recent evidence has argued against the existence of PKM isoform switch and related metabolic effects during cancer progression. We compared the effects of PKM1 and PKM2 in cell invasiveness and metastasis of pancreatic ductal adenocarcinoma (PDAC). Both PKM1 and PKM2 expression affected cell migration and invasion abilities of PDAC cells, but only knockdown of PKM2 suppressed metastasis in a xenograft model. By comparing the established PKM2 mutants in the regulation of cell invasion, we found that PKM2 may control cell mobility through its protein kinase and phopho-peptide binding abilities. Further survey for PKM2-associated proteins identified PAK2 as a possible phosphorylation target in PDAC. In vitro binding and kinase assays revealed that PKM2 directly phosphorylated PAK2 at Ser20, Ser141, and Ser192/197. Knockdown of PKM2 decreased PAK2 protein half-life by increasing ubiquitin-dependent proteasomal degradation. Moreover, we identified PAK2 as an HSP90 client protein and the mutation at Ser192/197 of PAK2 reduced PAK2–HSP90 association. Knockdown of PAK2 diminished in vitro cell mobility and in vivo metastatic ability of PKM2 overexpressed PDAC cells. PKM2 and PAK2 protein expression also positively correlated with each other in PDAC tissues. Our findings indicate that PKM2–PAK2 regulation is critical for developing metastasis in PDAC, and suggest that targeting the PKM2/HSP90/PAK2 complex has a potential therapeutic value in this deadly disease.

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Acknowledgements

We thank Dr. Zhi-Min Lu for the Flag-tagged K367M, K433E, and wild-type PKM2; Dr. Zhi-Ren Liu for HA-tagged R399E and wild-type PKM2. This work was supported by grants from the Ministry of Science and Technology of Taiwan, 104-0210-01-09-02, 105-0210-01-13-01, and 106-0210-01-15-02 (to M.H.); 105-2320-B-002-035-MY3 (to K.-T.H.); and 104-2314-B-002-027 (to T.-Y.C.).

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Affiliations

  1. Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan

    • Tsu-Yao Cheng
  2. Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan

    • Tsu-Yao Cheng
    •  & Hsiu-Po Wang
  3. Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan

    • Yi-Chieh Yang
  4. Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan

    • Yu-Wen Tien
  5. Department of Pathology, National Taiwan University Hospital, Taipei, Taiwan

    • Chia-Tung Shun
    •  & Hsin-Yi Huang
  6. The Genomics Research Center, Academia Sinica, Taipei, Taiwan

    • Michael Hsiao
  7. Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    • Michael Hsiao
  8. Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan

    • Kuo-Tai Hua

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The authors declare that they have no competing interests.

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Correspondence to Kuo-Tai Hua.

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https://doi.org/10.1038/s41388-017-0086-y

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