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RKTG inhibits angiogenesis by suppressing MAPK-mediated autocrine VEGF signaling and is downregulated in clear-cell renal cell carcinoma

Oncogene volume 29, pages 5404–5415 (2010)Cite this article

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Abstract

Vascular endothelial growth factors (VEGFs) are crucial regulators of angiogenesis and vasculogenesis. The autocrine VEGF signaling is required for maintaining the homeostasis of vasculature. Dysregulation of angiogenesis is implicated in the development of many human cancers, especially in clear-cell renal cell carcinoma (ccRCC), a highly vascularized tumor. Meanwhile, antiangiogenesis has become a mainstay in the treatment of human cancers. In this study, we analyzed the functional roles of RKTG (Raf Kinase Trapping to Golgi), a negative regulator of mitogen-activated protein kinase (Raf/MEK/ERK) signaling, by sequestration of Raf kinase to the Golgi apparatus, in angiogenesis and ccRCC. Through a series of in vitro and in vivo experiments, we found that RKTG has a negative effect on cell proliferation, migration, sprouting and angiogenesis of endothelial cells. RKTG, by suppressing mitogen-activated protein kinase signaling, negatively regulates the transactivation activity of hypoxia-inducible factor 1α (HIF-1α) by inhibiting formation of HIF-1α/p300 complex and suppressing VEGF transcription, thereby reducing hypoxia-induced VEGF production. The expression level of RKTG is significantly downregulated in clinical ccRCC tumor samples, with an inverse correlation with VEGF expression level. These results highlight the functional roles of RKTG and its regulated Raf/ERK/MEK signaling cascade in angiogenesis and autocrine VEGF signaling. In addition, this study indicates that RKTG is likely implicated in the development of ccRCC through its regulation on angiogenesis.

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Acknowledgements

We thank Dr George Simons and Dr Ilias Mylonis (University of Thessaly, Greece) for providing the plasmid of full-length HIF-1α fused to GST; Dr William E Carson III and Dr Gregory B Lesinski (Ohio State University, USA) for discussion about the CD31 immunohistochemistry experiment; Dr Young-Guen Kwon and Yong-Sun Maeng (Yonsei University, Korea) and Dr Morag Park and Caroline Saucier (McGill University Health Centre, Quebec, Canada) for discussion about the Matrigel plug assay experiment; Dr Luisa Iruela-Arispe and Dr Tom Chen (University of California, Los Angeles, USA) for providing the experimental protocols; and Dr Jurgen Seppen (Academic Medical Centre, Amsterdam, Netherlands) for discussion about the experiment with lentivirus. This work was supported by research grants from the Chinese Academy of Sciences (Knowledge Innovation Program KSCX1-YW-02), the National Natural Science Foundation of China (30830037) and the Ministry of Science and Technology of China (2007CB947100 and 2006CB943900) to YC. This work was also supported by the Ministry of Science and Technology of China (2010CB529506) to ZW.

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  1. Y Zhang and X Jiang: These authors contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai, China

    Y Zhang, X Jiang, W Liu, X Xie, Z Wang, J Fang & Y Chen

  2. Department of Urology, Fudan University Shanghai Cancer Center, Shanghai, China

    X Qin & D Ye

  3. Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China

    Z Yi & M Liu

  4. The First Hospital, Jilin University, Jilin, Changchun, China

    O Bai

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  1. Y Zhang
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Zhang, Y., Jiang, X., Qin, X. et al. RKTG inhibits angiogenesis by suppressing MAPK-mediated autocrine VEGF signaling and is downregulated in clear-cell renal cell carcinoma. Oncogene 29, 5404–5415 (2010). https://doi.org/10.1038/onc.2010.270

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