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Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth

Oncogene volume 20, pages 2727–2736 (2001)Cite this article

Abstract

In spite of the general recognition of von Hippel-Lindau (VHL) as a tumor suppressor gene, the physiological and pathological importance of VHL protein in cell growth regulation and tumorigenesis remains unclear. Here we show that in normal human renal proximal tubule epithelial cells (RPTEC), the steady-state amount of VHL protein is strictly regulated by cell density. The cellular VHL content is more than 100-fold higher in dense cultures than in sparse cultures. The increase in VHL protein at high cell density was also observed for NIH3T3 fibroblasts, suggesting the generality of the phenomenon. The growth rates of renal cell carcinoma cells lacking an intact VHL gene and their derivatives with wild-type or mutant VHL expression vector do not differ significantly when they are growing in log-phase. Importantly, however, there is a difference when they reach confluency: cells lacking wild-type VHL grew continuously, while cells expressing exogenous VHL protein showed relatively limited cell growth. Using an ecdysone-inducible VHL expressing cell line, we also show that the growth inhibition at high cell density can be released by attenuating the VHL expression. Taken together, we propose that VHL protein functions as a growth suppressor at high cell density, and this might be the basis of the tumor suppressor function of VHL.

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Abbreviations

RCC:

renal cell carcinoma

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

VEGF:

vascular endothelial growth factor

VHL:

von Hippel-Lindau

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Acknowledgements

We thank Dr William G Kaelin for 786-O cells and their VHL expressing subclones, and Dr Igor Kuzmin and Berton Zbar for UMRC-6 cells and their VHL expressing subclones, We thank Dr Teijiro Aso for anti elonginB antibody. We also thank Dr Kazuhiro Iwai for helpful discussion during the course of the work. This work was supported in part by Grants in Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan and Grants from the Japan Society for the Promotion of Science 12219215 (to S Ohno) and 11160216 (to S Hirai). M Baba received support from the Yokohama Foundation for Advancement of Medical Science.

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Authors and Affiliations

  1. Department of Molecular Biology, Yokohama City University School of Medicine, Yokohama, 236-0004, Japan

    Masaya Baba, Syu-ichi Hirai & Shigeo Ohno

  2. Department of Urology, Yokohama City University School of Medicine, Yokohama, 236-0004, Japan

    Masaya Baba, Satoshi Kawakami, Takeshi Kishida, Naoki Sakai, Shigeki Kaneko, Masahiro Yao, Yoshinobu Kubota & Masahiko Hosaka

  3. Department of Urology, Kochi Medical School, Kochi, 783-8505, Japan

    Taro Shuin

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  1. Masaya Baba
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Baba, M., Hirai, Si., Kawakami, S. et al. Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth. Oncogene 20, 2727–2736 (2001). https://doi.org/10.1038/sj.onc.1204397

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