Class II histone deacetylases play pivotal roles in heat shock protein 90-mediated proteasomal degradation of vascular endothelial growth factor receptors

https://doi.org/10.1016/j.bbrc.2008.01.056Get rights and content

Abstract

Vascular endothelial growth factor receptors (VEGFRs) perform pivotal roles in both tumor growth and angiogenesis. In this study, we report that histone deacetylase inhibitors (HDIs) induce a reduction in VEGFR1 and VEGFR2 protein expression via the inhibition of class II histone deacetylases (HDACs) in human cancer cell lines. After HDI treatment, VEGFR1 and VEGFR2 were shown to be downregulated in a proteasome-dependent manner. HDI treatment induced a reduction in the binding of heat shock protein (Hsp) 90 to VEGFR1 or VEGFR2, followed by an increase of the binding of Hsp70 to VEGFR1 or VEGFR2. However, we noted no remarkable changes in the binding of Hsp90/Hsp70 to VEGFR3. HDI treatment effectively inhibited the activities of HDAC6 and HDAC10. Furthermore, the knock-down of HDAC6 or HDAC10, which was accomplished via the siRNA transfection, induced depletion of VEGFR1 or VEGFR2 proteins. Overall, these results indicate that HDAC6 and HDAC10 play important roles in Hsp-mediated VEGFR regulation.

Section snippets

Materials and methods

Reagents. Antibody to HDAC6 (H-300) was obtained from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). Antibody to HDAC10 was purchased from Biovision Research Products (Mountain View, CA, USA). 17-AAG, an Hsp90 inhibitor, and antibodies to Hsp90 (SPA-830) and Hsp70/Hsc70 (SPA-820) were purchased from Stressgen (Victoria, British Columbia, Canada). Proteasomal inhibitors (MG132, Lactacystin), were obtained from Sigma (St. Louis, MO, USA). SAHA and LAQ824 were provided by In2Gen Co. (Seoul,

HDIs induced the depletion of VEGFR proteins in cancer cells

In order to assess the effects of HDI treatment on the expression of the VEGFR family, we initially examined the levels of VEGFR family protein (VEGFR-1, VEGFR-2, and VEGFR-3) expression in human lung cancer cells (HCT116) and human gastric cancer cells (SNU-620) following treatment with hydroxamate-based HDI, including SAHA and LAQ824. Interestingly, among these 3 proteins, VEGFR-1 and VEGFR-2 were determined to have been downregulated after SAHA treatment in a time-dependent manner in both

Acknowledgments

This study was supported in part by grants from Seoul National University Hospital (Grant #: 03-2006-012-0) and by the BK21 Project for Medicine, Dentistry, and pharmacy. We thank the members of Dr. Bang’s laboratory for helpful discussion.

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