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EGFL6 promotes breast cancer by simultaneously enhancing cancer cell metastasis and stimulating tumor angiogenesis

Oncogene volume 38, pages 2123–2134 (2019)Cite this article

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Abstract

EGFL6, a member of the EGF-like superfamily, plays an important role during embryonic development and has been implicated in promotion of tumor angiogenesis without affecting wound healing. There is very little known about the function of EGFL6 in cancer cells. Here, we investigated whether EGFL6 plays a direct role in cancer cells in addition to the promotion of tumor angiogenesis. Our study showed that EGFL6 promoted epithelial–mesenchymal transition (EMT) and stemness of breast cancer cells and increased cell migration and invasion in cell culture studies. We also found that EGFL6 reduced apoptotic signaling in cancer cells and promoted tumor growth in vivo. Importantly, expression of EGFL6 in cancer cells and tumor endothelial cells not only increased tumor angiogenesis but also promoted migration of cancer cells. Such dual engagement of cancer and stromal cells suggests crosstalk mediated by EGFL6 in the tumor microenvironment. Blockade of EGFL6 using our novel anti-EGFL6 monoclonal antibody significantly reduced cancer cell migration, tumor angiogenesis, and tumor growth in mouse xenograft tumor models. Silencing EGFL6 mRNA by shRNA transfection of cancer cells also significantly reduced cancer cell migration, tumor angiogenesis, and tumor growth in mouse xenograft tumor models. Taken together, the results of this study indicate that targeting EGFL6 is a unique strategy for inhibiting both cancer cell metastasis and tumor angiogenesis.

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References

  1. Yeung G, Mulero JJ, Berntsen RP, Loeb DB, Drmanac R, Ford JE. Cloning of a novel epidermal growth factor repeat containing gene EGFL6: expressed in tumor and fetal tissues. Genomics. 1999;62:304–7.

    Article  CAS  Google Scholar 

  2. Mas VR, Maluf DG, Archer KJ, Yanek KC, Fisher RA. Angiogenesis soluble factors as hepatocellular carcinoma noninvasive markers for monitoring hepatitis C virus cirrhotic patients awaiting liver transplantation. Transplantation. 2007;84:1262–71.

    Article  Google Scholar 

  3. Noh K, Mangala LS, Han HD, Zhang N, Pradeep S, Wu SY, et al. Differential effects of EGFL6 on tumor versus wound angiogenesis. Cell Rep. 2017;21:2785–95.

    Article  CAS  Google Scholar 

  4. Bai S, Ingram P, Chen YC, Deng N, Pearson A, Niknafs Y, et al. EGFL6 regulates the asymmetric division, maintenance, and metastasis of ALDH+ ovarian cancer cells. Cancer Res. 2016;76:6396–409.

    Article  CAS  Google Scholar 

  5. Larimer BM, Deutscher SL. Identification of a peptide from in vivo bacteriophage display with homology to EGFL6: a candidate tumor vasculature ligand in breast cancer. J Mol Biomark Diagn. 2014;5:178.

    PubMed  PubMed Central  Google Scholar 

  6. Chim SM, Qin A, Tickner J, Pavlos N, Davey T, Wang H, et al. EGFL6 promotes endothelial cell migration and angiogenesis through the activation of extracellular signal-regulated kinase. J Biol Chem. 2011;286:22035–46.

    Article  CAS  Google Scholar 

  7. Nichol D, Stuhlmann H. EGFL7: a unique angiogenic signaling factor in vascular development and disease. Blood. 2012;119:1345–52.

    Article  CAS  Google Scholar 

  8. Nichol D, Shawber C, Fitch MJ, Bambino K, Sharma A, Kitajewski J, et al. Impaired angiogenesis and altered Notch signaling in mice overexpressing endothelial Egfl7. Blood. 2010;116:6133–43.

    Article  CAS  Google Scholar 

  9. Garcia-Carbonero R, van Cutsem E, Rivera F, Jassem J, Gore I Jr, Tebbutt N, et al. Randomized phase II trial of parsatuzumab (anti-EGFL7) or placebo in combination with FOLFOX and bevacizumab for first-line metastatic colorectal cancer. oncologist. 2017;22:1281.

    Article  Google Scholar 

  10. von Pawel J, Spigel DR, Ervin T, Losonczy G, Barlesi F, Juhasz E, et al. Randomized phase II trial of parsatuzumab (anti-EGFL7) or placebo in combination with carboplatin, paclitaxel, and bevacizumab for first-line nonsquamous non-small cell lung cancer. Oncologist. 2018;23:654–e58.

    Article  Google Scholar 

  11. Lu C, Bonome T, Li Y, Kamat AA, Han LY, Schmandt R, et al. Gene alterations identified by expression profiling in tumor-associated endothelial cells from invasive ovarian carcinoma. Cancer Res. 2007;67:1757–68.

    Article  CAS  Google Scholar 

  12. Buckanovich RJ, Sasaroli D, O’Brien-Jenkins A, Botbyl J, Hammond R, Katsaros D, et al. Tumor vascular proteins as biomarkers in ovarian cancer. J Clin Oncol. 2007;25:852–61.

    Article  CAS  Google Scholar 

  13. Gyorffy B, Lanczky A, Eklund AC, Denkert C, Budczies J, Li Q, et al. An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients. Breast Cancer Res Treat. 2010;123:725–31.

    Article  Google Scholar 

  14. Brabletz T. EMT and MET in metastasis: where are the cancer stem cells?. Cancer Cell. 2012;22:699–701.

    Article  CAS  Google Scholar 

  15. Biddle A, Mackenzie IC. Cancer stem cells and EMT in carcinoma. Cancer Metastasis Rev. 2012;31:285–93.

    Article  Google Scholar 

  16. Wang X, Gong Y, Wang D, Xie Q, Zheng M, Zhou Y, et al. Analysis of gene expression profiling in meningioma: deregulated signaling pathways associated with meningioma and EGFL6 overexpression in benign meningioma tissue and serum. PLoS One. 2012;7:e52707.

    Article  CAS  Google Scholar 

  17. Wang Y, Shi J, Chai K, Ying X, Zhou BP. The role of Snail in EMT and tumorigenesis. Curr Cancer Drug Targets. 2013;13:963–72.

    Article  CAS  Google Scholar 

  18. Dong LJ, Hsieh JC, Chung AE. Two distinct cell attachment sites in entactin are revealed by amino acid substitutions and deletion of the RGD sequence in the cysteine-rich epidermal growth factor repeat 2. J Biol Chem. 1995;270:15838–43.

    Article  CAS  Google Scholar 

  19. Fujiwara H, Ferreira M, Donati G, Marciano DK, Linton JM, Sato Y, et al. The basement membrane of hair follicle stem cells is a muscle cell niche. Cell . 2011;144:577–89.

    Article  CAS  Google Scholar 

  20. Parker LH, Schmidt M, Jin SW, Gray AM, Beis D, Pham T, et al. The endothelial-cell-derived secreted factor Egfl7 regulates vascular tube formation. Nature. 2004;428:754–8.

    Article  CAS  Google Scholar 

  21. Gong C, Fang J, Li G, Liu HH, Liu ZS. Effects of microRNA-126 on cell proliferation, apoptosis and tumor angiogenesis via the down-regulating ERK signaling pathway by targeting EGFL7 in hepatocellular carcinoma. Oncotarget. 2017;8:52527–42.

    PubMed  PubMed Central  Google Scholar 

  22. Wang FY, Kang CS, Wang-Gou SY, Huang CH, Feng CY, Li XJ. EGFL7 is an intercellular EGFR signal messenger that plays an oncogenic role in glioma. Cancer Lett. 2017;384:9–18.

    Article  CAS  Google Scholar 

  23. Dudvarski Stankovic N, Bicker F, Keller S, Jones DT, Harter PN, Kienzle A, et al. EGFL7 enhances surface expression of integrin alpha5beta1 to promote angiogenesis in malignant brain tumors. EMBO Mol Med. 2018;10:e8420.

    Article  Google Scholar 

  24. Papaioannou D, Shen C, Nicolet D, McNeil B, Bill M, Karunasiri M, et al. Prognostic and biological significance of the proangiogenic factor EGFL7 in acute myeloid leukemia. Proc Natl Acad Sci USA. 2017;114:E4641–E7.

    Article  CAS  Google Scholar 

  25. Dhupkar P, Zhao H, Mujoo K, An Z, Zhang N. Crk II silencing down-regulates IGF-IR and inhibits migration and invasion of prostate cancer cells. Biochem Biophys Rep. 2016;8:382–8.

    PubMed  PubMed Central  Google Scholar 

  26. Salameh A, Fan X, Choi BK, Zhang S, Zhang N, An Z. HER3 and LINC00052 interplay promotes tumor growth in breast cancer. Oncotarget. 2017;8:6526–39.

    Article  Google Scholar 

  27. Fan X, Brezski RJ, Deng H, Dhupkar PM, Shi Y, Gonzalez A, et al. A novel therapeutic strategy to rescue the immune effector function of proteolytically inactivated cancer therapeutic antibodies. Mol Cancer Ther. 2015;14:681–91.

    Article  CAS  Google Scholar 

  28. Shi Y, Fan X, Meng W, Deng H, Zhang N, An Z. Engagement of immune effector cells by trastuzumab induces HER2/ERBB2 downregulation in cancer cells through STAT1 activation. Breast Cancer Res. 2014;16:R33.

    Article  Google Scholar 

  29. Zhang S, Mukherjee S, Fan X, Salameh A, Mujoo K, Huang Z, et al. Novel association of DJ-1 with HER3 potentiates HER3 activation and signaling in cancer. Oncotarget. 2016;7:65758–69.

    PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Dr. Wei Xiong and Ms. Hui Deng for their technical assistance in cell culture and antibody preparation. We also want to thank Dr. Yunfei Wen and Dr. Prahlad Ram for their suggestions and discussion on data analysis during the manuscript preparation. We thank Dr. Georgina Salazar for her critical editing of the manuscript. This study was supported in part by Welch Foundation grant (AU-0042-20030616) to ZA, and Cancer Prevention and Research Institute of Texas (CPRIT) Grants (RP150230 and RP150551) to ZA and NZ. This research was also supported by NIH support (P50 CA217685) to AKS, the Frank McGraw Memorial Chair in Cancer Research, and the ACS Research Professor Award.

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    Authors and Affiliations

    1. Jiangsu Institute of Clinical Immunology, The First Affiliated Hospital of Soochow University, 708 Renmin Road, Suzhou, China

      Jingnan An & Xue-Guang Zhang

    2. Texas Therapeutics Institute, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 1825 Pressler St., Houston, TX, 77030, USA

      Jingnan An, Yi Du, Xuejun Fan, Yanhong Wang, Zhiqiang An & Ningyan Zhang

    3. Department of Microbiology and Immunology, Shanxi Medical University, Taiyuan, Shanxi, China

      Yanhong Wang

    4. Department of Experimental Therapeutics, Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA

      Cristina Ivan

    5. Gynecologic Oncology & Reproductive Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA

      Anil K. Sood

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    An, J., Du, Y., Fan, X. et al. EGFL6 promotes breast cancer by simultaneously enhancing cancer cell metastasis and stimulating tumor angiogenesis. Oncogene 38, 2123–2134 (2019). https://doi.org/10.1038/s41388-018-0565-9

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