Abstract
Human melanomas show oncogenic B-Raf mutations, which activate the B-Raf/MKK/ERK cascade. We screened microarrays to identify cellular targets of this pathway, and found that genes upregulated by B-Raf/MKK/ERK showed highest association with cell-cycle regulators, whereas genes downregulated were most highly associated with axon guidance genes, including plexin–semaphorin family members. Plexin B1 was strongly inhibited by mitogen-activated protein kinase signaling in melanoma cells and melanocytes. In primary melanoma cells, plexin B1 blocked tumorigenesis as measured by growth of colonies in soft agar, spheroids in extracellular matrix and xenograft tumors. Tumor suppression depended on residues in the C-terminal domain of plexin B1, which mediate receptor GTPase activating protein activity, and also correlated with AKT inhibition. Interestingly, the inhibitory response to plexin B1 was reduced or absent in cells from a matched metastatic tumor, suggesting that changes occur in metastatic cells which bypass the tumor-suppressor mechanisms. Plexin B1 also inhibited cell migration, but this was seen in metastatic cells and not in matched primary cells. Thus, plexin B1 has tumor-suppressor function in early-stage cells, although suppressing migration in late-stage cells. Our findings suggest that B-Raf/MKK/ERK provides a permissive environment for melanoma genesis by modulating plexin B1.
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Acknowledgements
We thank Meenhard Herlyn for providing melanoma cell lines used in this study, and Silvio Gutkind, Kun-liang Guan, Xuedong Liu and Andrey Sorokin for expression plasmids. We also thank Norma Aumen for tumor sectioning, Helen Marshall for microarray analysis and Micah Hamady and Rob Knight for providing access to GO-Getter. This work was supported by NIH awards R01-CA118972 (NGA) and P50-CA058187 (DC), and fellowship awards F32-CA105796 (GMA), ACS-PF-04-152 (CHC) and T32-GM008759 (KLC).
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)
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Argast, G., Croy, C., Couts, K. et al. Plexin B1 is repressed by oncogenic B-Raf signaling and functions as a tumor suppressor in melanoma cells. Oncogene 28, 2697–2709 (2009). https://doi.org/10.1038/onc.2009.133
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DOI: https://doi.org/10.1038/onc.2009.133
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