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Transferrin receptor 1 promotes the fibroblast growth factor receptor-mediated oncogenic potential of diffused-type gastric cancer

Oncogene volume 41, pages 2587–2596 (2022)Cite this article

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Abstract

Diffuse-type gastric cancer (DGC) is a highly invasive subtype of gastric adenocarcinoma that frequently exhibits scattered peritoneal metastasis. Previous studies have shown that the genes of receptor tyrosine kinases (RTKs), such as fibroblast growth factor receptor 2 (FGFR2) or Met, are amplified in some DGC cell lines, leading to the constitutive activation of corresponding RTKs. In these cell lines, the survival of cancer cells appears to be dependent on the activation of RTKs. To gain novel insights into the downstream signaling pathways of RTKs specific to DGC, phosphotyrosine-containing proteins associated with activated FGFR2 were purified through two sequential rounds of immunoprecipitation from the lysates of two DGC cell lines. As a result, transferrin receptor 1 (TfR1) was identified as the binding partner of FGFR2. Biochemical analysis confirmed that TfR1 protein binds to FGFR2 and is phosphorylated at tyrosine 20 (Tyr20) in an FGFR2 kinase activity-dependent manner. The knockdown of TfR1 and treatment with an inhibitor of FGFR2 caused significant impairment in iron uptake and suppression of cellular proliferation in vitro. Moreover, the suppression of expression levels of TfR1 in the DGC cells significantly reduced their tumorigenicity and potency of peritoneal dissemination. It was indicated that TfR1, when phosphorylated by the binding partner FGFR2 in DGC cells, promotes proliferation and tumorigenicity of these cancer cells. These results suggest that the control of TfR1 function may serve as a therapeutic target in DGC with activated FGFR2.

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Fig. 1: Identification of FGFR2-binding phospho-target proteins in DGC cell lines.
Fig. 2: Tyrosine phosphorylation of TfR1 and its association with FGFR2.
Fig. 3: Requirement of FGFR2-mediated Tyr20 phosphorylation of TfR1 for transferrin internalization.
Fig. 4: Iron uptake and cell proliferation by phospho-TfR1 specific for FGFR-type DGC cell lines.
Fig. 5: Effect of TfR1 knockdown on tumor growth and peritoneal metastasis in the mouse model of intraperitoneal transplant.
Fig. 6: The regulation of cell proliferation and survival mediated by TfR1 in FGFR-type DGC cells.

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Acknowledgements

We thank Dr. Kazuyoshi Yanagihara (National Cancer Center, Japan) for providing 44As3 and 58As9 cell lines. This work was supported by JSPS KAKENHI Grants Numbers JP17K08648, JP18K07242, and JP21K07105.

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Authors and Affiliations

  1. Department of Biochemistry, Kitasato University School of Medicine, Sagamihara, Japan

    Takuya Shirakihara & Ryuichi Sakai

  2. Department of Cancer Cell Research, Sasaki Institute, Sasaki Foundation, Tokyo, Japan

    Hideki Yamaguchi

  3. Division of Rare Cancer Research, National Cancer Center Research Institute, Tokyo, Japan

    Tadashi Kondo

  4. Molecular Oncology and Therapeutics, Osaka City University Graduate School of Medicine, Osaka, Japan

    Masakazu Yashiro

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Contributions

Study conception and design: TS, RS. Data acquisition: TS, TK. Data analysis and interpretation: TS, HY, RS. Resources: MY. Paper preparation: TS, RS. Paper review & editing: HY, TK, MY.

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Correspondence to Ryuichi Sakai.

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Shirakihara, T., Yamaguchi, H., Kondo, T. et al. Transferrin receptor 1 promotes the fibroblast growth factor receptor-mediated oncogenic potential of diffused-type gastric cancer. Oncogene 41, 2587–2596 (2022). https://doi.org/10.1038/s41388-022-02270-5

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