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PRRG4 promotes breast cancer metastasis through the recruitment of NEDD4 and downregulation of Robo1

Oncogene volume 39, pages 7196–7208 (2020)Cite this article

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Abstract

Metastasis is responsible for the death of most breast cancer patients. Robo1 has been implicated as a tumor suppressor for various cancers including breast cancer. However, it is not well understood how Robo1 expression is regulated during tumorigenesis. In this study, we uncovered that the transmembrane proline rich γ-carboxyglutamic acid protein 4 (PRRG4) promotes breast cancer metastasis by downregulating Robo1. Analysis of mRNA expression data in The Cancer Genome Atlas and immunohistochemistry assay on breast tumor samples showed that PRRG4 expression was higher in breast tumors than in normal breast tissues. Experiments with PRRG4 knockdown and overexpression revealed that PRRG4 promoted migration and invasion of breast cancer cells, and enhanced metastasis in an experimental metastasis model. Mechanistically, we found that PRRG4 via its LPSY and PPPY motifs recruited the E3 ubiquitin ligase NEDD4, which induced ubiquitination and degradation of Robo1, thus contributing to migration and invasion of breast cancer cells. In addition, PRRG4 interacted with and enhanced protein tyrosine kinase Src and FAK activation. Overall, our data support a model that PRRG4 via NEDD4 downregulates the Robo1, resulting in the activation of Src and FAK and promoting breast cancer metastasis. PRRG4 may be a novel target for treating metastatic breast cancer.

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Fig. 1: PRRG4 expression is higher in breast tumors than in normal breast tissues.
Fig. 2: PRRG4 promotes migration and invasion of breast cancer cells.
Fig. 3: PRRG4 downregulates Robo1 by increasing Robo1 protein turnover in breast cancer cells.
Fig. 4: PRRG4 downregulates Robo1 by recruiting NEDD4 and promoting Robo1 ubiquitination in breast cancer cells.
Fig. 5: PRRG4 promotes migration and invasion of breast cancer cells through NEDD4 and downregulation of Robo1.
Fig. 6: PRRG4 promotes Src and FAK activation via downregulation of the Robo1.
Fig. 7: PRRG4 via its WW domain binding motif promotes breast cancer metastasis.
Fig. 8: A working model of PRRG4 promoting metastasis of breast cancer cells.

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Acknowledgements

We thank Dr. Weibao Jiao (Kunming Institute of Zoology, Academia Sinica) for kindly providing us the MDA-MB-231-luciferase cells, and Helen Gu (Department of Economics, Massachusetts Institute of Technology) for proofreading the paper. This work was supported by grants (81972463 and 81772819) from the National Natural Science Foundation of China, and grants (LY20C080001 and LY18H080003) from Natural Science Foundation of Zhejiang Province, China, and was supported in part by Wenzhou Medical University Startup funds and the Key Discipline of Zhejiang Province in Medical Technology (First Class, Category A).

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  1. These authors contributed equally: Lingling Zhang, Yaqian Qin

Authors and Affiliations

  1. Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, China

    Lingling Zhang, Yaqian Qin, Guang Wu, Jieyi Wang, Jiawei Cao, Yaqi Wang, Du Wu, Licai He, Jianxin Lyu, Hongzhi Li & Haihua Gu

  2. Department of Pathology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325035, China

    Kaiyan Yang

  3. Department of Pathology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, China

    Zhiguang Zhao

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Zhang, L., Qin, Y., Wu, G. et al. PRRG4 promotes breast cancer metastasis through the recruitment of NEDD4 and downregulation of Robo1. Oncogene 39, 7196–7208 (2020). https://doi.org/10.1038/s41388-020-01494-7

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