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A novel and selective inhibitor of PKC ζ potently inhibits human breast cancer metastasis in vitro and in mice

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Tumor Biology

Abstract

Cell motility and chemotaxis play pivotal roles in the process of tumor development and metastasis. Protein kinase C ζ (PKC ζ) mediates epidermal growth factor (EGF)-stimulated chemotactic signaling pathway through regulating cytoskeleton rearrangement and cell adhesion. The purpose of this study was to develop anti-PKC ζ therapeutics for breast cancer metastasis. In this study, a novel and high-efficient PKC ζ inhibitor named PKCZI195.17 was screened out through a substrate-specific strategy. MTT assay was used to determine the cell viability of human breast cancer MDA-MB-231, MDA-MB-435, and MCF-7 cells while under PKCZI195.17 treatment. Wound-healing, chemotaxis, and Matrigel invasion assays were performed to detect the effects of PKCZI195.17 on breast cancer cells migration and invasion. Adhesion, actin polymerization, and Western blotting were performed to detect the effects of PKCZI195.17 on cells adhesion and actin polymerization, and explore the downsteam signaling mechanisms involved in PKC ζ inhibition. MDA-MB-231 xenograft was used to measure the in vivo anti-metastasis efficacy of PKCZI195.17. The compound PKCZI195.17 selectively inhibited PKC ζ kinase activity since it failed to inhibit PKC α, PKC β, PKC δ, PKC η, AKT2, as well as FGFR2 activity. PKCZI195.17 significantly impaired spontaneous migration, chemotaxis, and invasion of human breast cancer MDA-MB-231, MDA-MB-435, and MCF-7 cells, while PKCZI195.17 did not obviously inhibited cells viability. PKCZI195.17 also inhibited cells adhesion and actin polymerization through attenuating the phosphorylations of integrin β1, LIMK, and cofilin, which might be the downstream effectors of PKC ζ-mediated chemotaxis in MDA-MB-231 cells. Furthermore, PKCZI195.17 suppressed the breast cancer metastasis and increased the survival time of breast tumor-bearing mice. In summary, PKCZI195.17 was a PKC ζ-specific inhibitor which dampened cancer cell migration and metastasis and may serve as a novel therapeutic drug for breast cancer metastasis.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81301985 and No. 81102292) and the Natural Science Foundation of Tianjin (No. 14JCQNJC14000 and No. 12JCQNJC06200; to Jing Wu and Rui Yang).

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Authors and Affiliations

  1. Department of Clinical Laboratory, Tianjin Third Central Hospital, No. 83 Jintang Road, Hedong District, Tianjin, 300170, China

    Jing Wu, Shuye Liu, Zhijuan Fan, Lei Zhang & Yaqiong Tian

  2. Research Center of Basic Medical Science, Tianjin Medical University, No. 22 Qixiangtai Road, Heping District, Tianjin, 300070, China

    Rui Yang

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  1. Jing Wu
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  2. Shuye Liu
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  3. Zhijuan Fan
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  4. Lei Zhang
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  5. Yaqiong Tian
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Correspondence to Jing Wu or Rui Yang.

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All animal experiments were conducted in accordance with the Guide for the Care and Use of Laboratory Animal of Tianjin Third Central Hospital.

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Wu, J., Liu, S., Fan, Z. et al. A novel and selective inhibitor of PKC ζ potently inhibits human breast cancer metastasis in vitro and in mice. Tumor Biol. 37, 8391–8401 (2016). https://doi.org/10.1007/s13277-015-4744-9

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  • DOI: https://doi.org/10.1007/s13277-015-4744-9

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