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Downregulation of ROBO4 in Pancreatic Cancer Serves as a Biomarker of Poor Prognosis and Indicates Increased Cell Motility and Proliferation Through Activation of MMP-9

  • Translational Research
  • Published:
Annals of Surgical Oncology Aims and scope Submit manuscript

Abstract

Background

The axon guidance gene family, SLIT/ROBO pathway, controls neural network formation, which correlates with the development of several cancers.

Methods

We found through analysis of the public database that ROBO4, one of the axon guidance molecules among the SLIT/ROBO family, is significantly downregulated in primary pancreatic cancer tissues compared with adjacent normal tissues. We carried out transfection experiments using three pancreatic cancer cell lines (MiaPaCa-2, BxPC-3, and SW1990) and one pancreatic duct epithelial cell line (HPDE6c7). A total of 51 clinical samples were then examined by immunohistochemical staining to find an association between ROBO4 expression at the protein level, clinical characteristics, and surgical outcomes.

Results

ROBO4 overexpression suppressed the invasion and migration abilities in MiaPaCa-2 and BxPC-3, while ROBO4 siRNA transfection to SW1990 and HPDE6c7 enhanced those activities. PCR-based profiling detected MMP-9 as a candidate downstream target of ROBO4, which was validated by decreased MMP-9 activity after the ROBO4 overexpression assay. High ROBO4 expression clinical samples had significantly better overall survival rather than low ROBO4 cases (P = 0.048).

Conclusion

These findings suggest that decreased ROBO4 expression activates malignant phenotypes in cancer cells and is correlated with poor survival outcomes in pancreatic cancer.

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Acknowledgement

We thank James P. Mahaffey, PhD, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

Funding

This study was supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant-in-Aid for Young Scientists, JP19K18080.

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Authors and Affiliations

  1. Department of Gastroenterological Surgery (Surgery II), Nagoya University Graduate School of Medicine, Nagoya, Japan

    Masaya Yamanaka MD, Masamichi Hayashi MD, PhD, Fuminori Sonohara MD, PhD, Suguru Yamada MD, PhD, Haruyoshi Tanaka MD, PhD, Daigo Kobayashi MD, Keisuke Kurimoto MD, PhD, Nobutake Tanaka MD, PhD, Yoshikuni Inokawa MD, PhD, Hideki Takami MD, PhD, Norifumi Hattori MD, PhD, Mitsuro Kanda MD, PhD, Chie Tanaka MD, PhD, Goro Nakayama MD, PhD, Masahiko Koike MD, PhD & Yasuhiro Kodera MD, PhD

  2. Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan

    Akihiro Sakai MD & Shinji Mii MD, PhD

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  1. Masaya Yamanaka MD
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Correspondence to Masamichi Hayashi MD, PhD.

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Yamanaka, M., Hayashi, M., Sonohara, F. et al. Downregulation of ROBO4 in Pancreatic Cancer Serves as a Biomarker of Poor Prognosis and Indicates Increased Cell Motility and Proliferation Through Activation of MMP-9. Ann Surg Oncol 29, 7180–7189 (2022). https://doi.org/10.1245/s10434-022-12039-5

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  • DOI: https://doi.org/10.1245/s10434-022-12039-5

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