The RASSF1A Tumor Suppressor Binds the RasGAP DAB2IP and Modulates RAS Activation in Lung Cancer
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. RASSF1A Binds the RASGAP DAB2IP
2.2. RASSF1A Modulates DAB2IP Expression
2.3. RASSF1A May Act with DAB2IP to Regulate RAS Activation
2.4. Dual Inhibition of RASSF1A and DAB2IP Has a Synergistic Effect on Growth and Transformation of a Wild-Type RAS Cell Line
3. Discussion
4. Materials and Methods
4.1. Plasmids and shRNAs
4.2. Tissue Culture and Cell Lines
4.3. D Growth Assay
4.4. Xenograft Growth Assay
4.5. Pulldown Assays and Western Blot Analysis
4.6. Fluorescence Microscopy
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Prior, I.A.; Lewis, P.D.; Mattos, C. A comprehensive survey of Ras mutations in cancer. Cancer Res. 2012, 72, 2457–2467. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Murugan, A.K.; Grieco, M.; Tsuchida, N. RAS mutations in human cancers: Roles in precision medicine. Semin. Cancer Biol. 2019, 59, 23–35. [Google Scholar] [CrossRef] [PubMed]
- Hurley, J.B.; Simon, M.I.; Teplow, D.B.; Robishaw, J.D.; Gilman, A.G. Homologies between signal transducing G proteins and ras gene products. Science 1984, 226, 860–862. [Google Scholar] [CrossRef] [PubMed]
- Bos, J.L.; Rehmann, H.; Wittinghofer, A. GEFs and GAPs: Critical elements in the control of small G proteins. Cell 2007, 129, 865–877. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhang, F.; Cheong, J.K. The renewed battle against RAS-mutant cancers. Cell. Mol. Life Sci. CMLS 2016, 73, 1845–1858. [Google Scholar] [CrossRef] [PubMed]
- Vigil, D.; Cherfils, J.; Rossman, K.L.; Der, C.J. Ras superfamily GEFs and GAPs: Validated and tractable targets for cancer therapy? Nat. Rev. Cancer 2010, 10, 842–857. [Google Scholar] [CrossRef] [Green Version]
- Tartaglia, M.; Pennacchio, L.A.; Zhao, C.; Yadav, K.K.; Fodale, V.; Sarkozy, A.; Pandit, B.; Oishi, K.; Martinelli, S.; Schackwitz, W.; et al. Gain-of-function SOS1 mutations cause a distinctive form of Noonan syndrome. Nat. Genet. 2007, 39, 75–79. [Google Scholar] [CrossRef]
- Wee, P.; Wang, Z. Epidermal Growth Factor Receptor Cell Proliferation Signaling Pathways. Cancers 2017, 9, 52. [Google Scholar] [CrossRef] [Green Version]
- Maertens, O.; Cichowski, K. An expanding role for RAS GTPase activating proteins (RAS GAPs) in cancer. Adv. Biol. Regul. 2014, 55, 1–14. [Google Scholar] [CrossRef]
- McCormick, F. Going for the GAP. Curr. Biol. 1998, 8, R673–R674. [Google Scholar] [CrossRef] [Green Version]
- O’Bryan, J.P. Pharmacological targeting of RAS: Recent success with direct inhibitors. Pharmacol. Res. 2019, 139, 503–511. [Google Scholar] [CrossRef] [PubMed]
- Cox, A.D.; Der, C.J. The dark side of Ras: Regulation of apoptosis. Oncogene 2003, 22, 8999–9006. [Google Scholar] [CrossRef] [PubMed]
- Donninger, H.; Schmidt, M.L.; Mezzanotte, J.; Barnoud, T.; Clark, G.J. Ras signaling through RASSF proteins. Semin. Cell Dev. Biol. 2016, 58, 86–95. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fausti, F.; Di Agostino, S.; Sacconi, A.; Strano, S.; Blandino, G. Hippo and rassf1a Pathways: A Growing Affair. Mol. Biol. Int. 2012, 2012, 307628. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Matallanas, D.; Romano, D.; Al-Mulla, F.; O’Neill, E.; Al-Ali, W.; Crespo, P.; Doyle, B.; Nixon, C.; Sansom, O.; Drosten, M.; et al. Mutant K-Ras activation of the proapoptotic MST2 pathway is antagonized by wild-type K-Ras. Mol. Cell 2011, 44, 893–906. [Google Scholar] [CrossRef] [Green Version]
- Vos, M.D.; Dallol, A.; Eckfeld, K.; Allen, N.P.; Donninger, H.; Hesson, L.B.; Calvisi, D.; Latif, F.; Clark, G.J. The RASSF1A tumor suppressor activates Bax via MOAP-1. J. Biol. Chem. 2006, 281, 4557–4563. [Google Scholar] [CrossRef] [Green Version]
- Hesson, L.B.; Cooper, W.N.; Latif, F. The role of RASSF1A methylation in cancer. Dis. Markers 2007, 23, 73–87. [Google Scholar] [CrossRef] [Green Version]
- Avruch, J.; Praskova, M.; Ortiz-Vega, S.; Liu, M.; Zhang, X.F. Nore1 and RASSF1 regulation of cell proliferation and of the MST1/2 kinases. Methods Enzymol. 2006, 407, 290–310. [Google Scholar] [CrossRef]
- Dubois, F.; Keller, M.; Calvayrac, O.; Soncin, F.; Hoa, L.; Hergovich, A.; Parrini, M.C.; Mazieres, J.; Vaisse-Lesteven, M.; Camonis, J.; et al. RASSF1A Suppresses the Invasion and Metastatic Potential of Human Non-Small Cell Lung Cancer Cells by Inhibiting YAP Activation through the GEF-H1/RhoB Pathway. Cancer Res. 2016, 76, 1627–1640. [Google Scholar] [CrossRef] [Green Version]
- Chamberlain, C.E.; Scheel, D.W.; McGlynn, K.; Kim, H.; Miyatsuka, T.; Wang, J.; Nguyen, V.; Zhao, S.; Mavropoulos, A.; Abraham, A.G.; et al. Menin determines K-RAS proliferative outputs in endocrine cells. J. Clin. Investig. 2014, 124, 4093–4101. [Google Scholar] [CrossRef] [Green Version]
- Schmidt, M.L.; Hobbing, K.R.; Donninger, H.; Clark, G.J. RASSF1A Deficiency Enhances RAS-Driven Lung Tumorigenesis. Cancer Res. 2018, 78, 2614–2623. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kim, D.H.; Kim, J.S.; Park, J.H.; Lee, S.K.; Ji, Y.I.; Kwon, Y.M.; Shim, Y.M.; Han, J.; Park, J. Relationship of Ras association domain family 1 methylation and K-ras mutation in primary non-small cell lung cancer. Cancer Res. 2003, 63, 6206–6211. [Google Scholar] [PubMed]
- Thaler, S.; Hahnel, P.S.; Schad, A.; Dammann, R.; Schuler, M. RASSF1A mediates p21Cip1/Waf1-dependent cell cycle arrest and senescence through modulation of the Raf-MEK-ERK pathway and inhibition of Akt. Cancer Res. 2009, 69, 1748–1757. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Li, W.; Yue, F.; Dai, Y.; Shi, B.; Xu, G.; Jiang, X.; Zhou, X.; Pfeifer, G.P.; Liu, L. Suppressor of hepatocellular carcinoma RASSF1A activates autophagy initiation and maturation. Cell Death Differ. 2019, 26, 1379–1395. [Google Scholar] [CrossRef] [Green Version]
- Romano, D.; Nguyen, L.K.; Matallanas, D.; Halasz, M.; Doherty, C.; Kholodenko, B.N.; Kolch, W. Protein interaction switches coordinate Raf-1 and MST2/Hippo signalling. Nat. Cell Biol. 2014, 16, 673–684. [Google Scholar] [CrossRef]
- Kilili, G.K.; Kyriakis, J.M. Mammalian Ste20-like kinase (Mst2) indirectly supports Raf-1/ERK pathway activity via maintenance of protein phosphatase-2A catalytic subunit levels and consequent suppression of inhibitory Raf-1 phosphorylation. J. Biol. Chem. 2010, 285, 15076–15087. [Google Scholar] [CrossRef] [Green Version]
- Zhang, H.; He, Y.; Dai, S.; Xu, Z.; Luo, Y.; Wan, T.; Luo, D.; Jones, D.; Tang, S.; Chen, H.; et al. AIP1 functions as an endogenous inhibitor of VEGFR2-mediated signaling and inflammatory angiogenesis in mice. J. Clin. Investig. 2008, 118, 3904–3916. [Google Scholar] [CrossRef] [Green Version]
- Huang, Q.; Qin, L.; Dai, S.; Zhang, H.; Pasula, S.; Zhou, H.; Chen, H.; Min, W. AIP1 suppresses atherosclerosis by limiting hyperlipidemia-induced inflammation and vascular endothelial dysfunction. Arterioscler. Thromb. Vasc. Biol. 2013, 33, 795–804. [Google Scholar] [CrossRef] [Green Version]
- Xie, D.; Gore, C.; Liu, J.; Pong, R.C.; Mason, R.; Hao, G.; Long, M.; Kabbani, W.; Yu, L.; Zhang, H.; et al. Role of DAB2IP in modulating epithelial-to-mesenchymal transition and prostate cancer metastasis. Proc. Natl. Acad. Sci. USA 2010, 107, 2485–2490. [Google Scholar] [CrossRef] [Green Version]
- Sun, L.; Yao, Y.; Lu, T.; Shang, Z.; Zhan, S.; Shi, W.; Pan, G.; Zhu, X.; He, S. DAB2IP Downregulation Enhances the Proliferation and Metastasis of Human Gastric Cancer Cells by Derepressing the ERK1/2 Pathway. Gastroenterol. Res. Pract. 2018, 2018, 2968252. [Google Scholar] [CrossRef]
- Wang, Z.; Tseng, C.P.; Pong, R.C.; Chen, H.; McConnell, J.D.; Navone, N.; Hsieh, J.T. The mechanism of growth-inhibitory effect of DOC-2/DAB2 in prostate cancer. Characterization of a novel GTPase-activating protein associated with N-terminal domain of DOC-2/DAB2. J. Biol. Chem. 2002, 277, 12622–12631. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vos, M.D.; Martinez, A.; Elam, C.; Dallol, A.; Taylor, B.J.; Latif, F.; Clark, G.J. A role for the RASSF1A tumor suppressor in the regulation of tubulin polymerization and genomic stability. Cancer Res. 2004, 64, 4244–4250. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dallol, A.; Agathanggelou, A.; Fenton, S.L.; Ahmed-Choudhury, J.; Hesson, L.; Vos, M.D.; Clark, G.J.; Downward, J.; Maher, E.R.; Latif, F. RASSF1A interacts with microtubule-associated proteins and modulates microtubule dynamics. Cancer Res. 2004, 64, 4112–4116. [Google Scholar] [CrossRef] [Green Version]
- Liu, L.; Tommasi, S.; Lee, D.H.; Dammann, R.; Pfeifer, G.P. Control of microtubule stability by the RASSF1A tumor suppressor. Oncogene 2003, 22, 8125–8136. [Google Scholar] [CrossRef] [Green Version]
- Huang, Y.Z.; Wu, W.; Wu, K.; Xu, X.N.; Tang, W.R. Association of RASSF1A promoter methylation with lung cancer risk: A meta-analysis. Asian Pac. J. Cancer Prev. 2014, 15, 10325–10328. [Google Scholar] [CrossRef] [PubMed]
- Dote, H.; Toyooka, S.; Tsukuda, K.; Yano, M.; Ota, T.; Murakami, M.; Naito, M.; Toyota, M.; Gazdar, A.F.; Shimizu, N. Aberrant promoter methylation in human DAB2 interactive protein (hDAB2IP) gene in gastrointestinal tumour. Br. J. Cancer 2005, 92, 1117–1125. [Google Scholar] [CrossRef] [Green Version]
- Lito, P.; Solomon, M.; Li, L.S.; Hansen, R.; Rosen, N. Allele-specific inhibitors inactivate mutant KRAS G12C by a trapping mechanism. Science 2016, 351, 604–608. [Google Scholar] [CrossRef] [Green Version]
- Vicent, S.; Chen, R.; Sayles, L.C.; Lin, C.; Walker, R.G.; Gillespie, A.K.; Subramanian, A.; Hinkle, G.; Yang, X.; Saif, S.; et al. Wilms tumor 1 (WT1) regulates KRAS-driven oncogenesis and senescence in mouse and human models. J. Clin. Investig. 2010, 120, 3940–3952. [Google Scholar] [CrossRef]
- Hofmann, I.; Weiss, A.; Elain, G.; Schwaederle, M.; Sterker, D.; Romanet, V.; Schmelzle, T.; Lai, A.; Brachmann, S.M.; Bentires-Alj, M.; et al. K-RAS mutant pancreatic tumors show higher sensitivity to MEK than to PI3K inhibition in vivo. PLoS ONE 2012, 7, e44146. [Google Scholar] [CrossRef] [Green Version]
- Donninger, H.; Clark, J.A.; Monaghan, M.K.; Schmidt, M.L.; Vos, M.; Clark, G.J. Cell cycle restriction is more important than apoptosis induction for RASSF1A protein tumor suppression. J. Biol. Chem. 2014, 289, 31287–31295. [Google Scholar] [CrossRef] [Green Version]
- Shivakumar, L.; Minna, J.; Sakamaki, T.; Pestell, R.; White, M.A. The RASSF1A tumor suppressor blocks cell cycle progression and inhibits cyclin D1 accumulation. Mol. Cell Biol. 2002, 22, 4309–4318. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tartaglia, M.; Gelb, B.D. Disorders of dysregulated signal traffic through the RAS-MAPK pathway: Phenotypic spectrum and molecular mechanisms. Ann. N. Y. Acad. Sci. 2010, 1214, 99–121. [Google Scholar] [CrossRef] [Green Version]
- Malumbres, M.; Barbacid, M. RAS oncogenes: The first 30 years. Nat. Rev. Cancer 2003, 3, 459–465. [Google Scholar] [CrossRef] [PubMed]
- Donninger, H.; Vos, M.D.; Clark, G.J. The RASSF1A tumor suppressor. J. Cell Sci. 2007, 120, 3163–3172. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Harrell Stewart, D.R.; Clark, G.J. Pumping the brakes on RAS-negative regulators and death effectors of RAS. J. Cell Sci. 2020, 133. [Google Scholar] [CrossRef] [PubMed]
- Agathanggelou, A.; Bieche, I.; Ahmed-Choudhury, J.; Nicke, B.; Dammann, R.; Baksh, S.; Gao, B.; Minna, J.D.; Downward, J.; Maher, E.R.; et al. Identification of novel gene expression targets for the Ras association domain family 1 (RASSF1A) tumor suppressor gene in non-small cell lung cancer and neuroblastoma. Cancer Res. 2003, 63, 5344–5351. [Google Scholar] [PubMed]
- Dubois, F.; Bergot, E.; Zalcman, G.; Levallet, G. RASSF1A, puppeteer of cellular homeostasis, fights tumorigenesis, and metastasis-an updated review. Cell Death Dis. 2019, 10, 928. [Google Scholar] [CrossRef]
- Yu, L.; Shang, Z.F.; Abdisalaam, S.; Lee, K.J.; Gupta, A.; Hsieh, J.T.; Asaithamby, A.; Chen, B.P.; Saha, D. Tumor suppressor protein DAB2IP participates in chromosomal stability maintenance through activating spindle assembly checkpoint and stabilizing kinetochore-microtubule attachments. Nucleic Acids Res. 2016, 44, 8842–8854. [Google Scholar] [CrossRef] [Green Version]
- Jackson, P.K. Linking tumor suppression, DNA damage and the anaphase-promoting complex. Trends Cell Biol. 2004, 14, 331–334. [Google Scholar] [CrossRef]
- Patricelli, M.P.; Janes, M.R.; Li, L.S.; Hansen, R.; Peters, U.; Kessler, L.V.; Chen, Y.; Kucharski, J.M.; Feng, J.; Ely, T.; et al. Selective Inhibition of Oncogenic KRAS Output with Small Molecules Targeting the Inactive State. Cancer Discov. 2016, 6, 316–329. [Google Scholar] [CrossRef] [Green Version]
- Kessler, D.; Gmachl, M.; Mantoulidis, A.; Martin, L.J.; Zoephel, A.; Mayer, M.; Gollner, A.; Covini, D.; Fischer, S.; Gerstberger, T.; et al. Drugging an undruggable pocket on KRAS. Proc. Natl. Acad. Sci. USA 2019, 116, 15823–15829. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhang, Y.; Meng, X.; Tang, H.; Cheng, M.; Yang, F.; Xu, W. Design, synthesis, and biological evaluation of novel substituted thiourea derivatives as potential anticancer agents for NSCLC by blocking K-Ras protein-effectors interactions. J. Enzym. Inhib. Med. Chem. 2020, 35, 344–353. [Google Scholar] [CrossRef] [PubMed]
- Donninger, H.; Clark, J.; Rinaldo, F.; Nelson, N.; Barnoud, T.; Schmidt, M.L.; Hobbing, K.R.; Vos, M.D.; Sils, B.; Clark, G.J. The RASSF1A tumor suppressor regulates XPA-mediated DNA repair. Mol. Cell Biol. 2015, 35, 277–287. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lai, C.H.; Chang, C.S.; Liu, H.H.; Tsai, Y.S.; Hsu, F.M.; Yu, Y.L.; Lai, C.K.; Gandee, L.; Pong, R.C.; Hsu, H.W.; et al. Sensitization of radio-resistant prostate cancer cells with a unique cytolethal distending toxin. Oncotarget 2014, 5, 5523–5534. [Google Scholar] [CrossRef] [Green Version]
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Harrell Stewart, D.R.; Schmidt, M.L.; Donninger, H.; Clark, G.J. The RASSF1A Tumor Suppressor Binds the RasGAP DAB2IP and Modulates RAS Activation in Lung Cancer. Cancers 2020, 12, 3807. https://doi.org/10.3390/cancers12123807
Harrell Stewart DR, Schmidt ML, Donninger H, Clark GJ. The RASSF1A Tumor Suppressor Binds the RasGAP DAB2IP and Modulates RAS Activation in Lung Cancer. Cancers. 2020; 12(12):3807. https://doi.org/10.3390/cancers12123807
Chicago/Turabian StyleHarrell Stewart, Desmond R., M. Lee Schmidt, Howard Donninger, and Geoffrey J. Clark. 2020. "The RASSF1A Tumor Suppressor Binds the RasGAP DAB2IP and Modulates RAS Activation in Lung Cancer" Cancers 12, no. 12: 3807. https://doi.org/10.3390/cancers12123807