Abstract
Small GTPases, particularly the Rho family, are key regulators of cell motility and migration. Dock180 was well known for the main target of signal adaptor protein Crk and acted as a guanine-nucleotide exchange factor for small GTPase Rac1. In the present study, Dock180 was found to combine primarily with CrkI other than CrkII, and its association with Elmo1 was also demonstrated in ovarian cancer cell SKOV3. To evaluate the role of Dock180 in human ovarian cancer cell, we performed RNAi-mediated knockdown of Dock180 in SKOV3 cells using small interfering RNA expression vector. In Dock180 knockdown cells, we found that Elmo1 expression and Rac1 activity were decreased simultaneously. By contrast, the expressions of both another Crk-combining molecule C3G and Rap1 activity were observed to increase obviously. Accordingly, all Dock180 knockdown cells present with evident change in cell morphology, reduced cell proliferation, and attenuated cell migration. Taken together, these results suggest that signal transfer of Crk/Dock180/Rac1 is implicated in actin cytoskeleton reorganization and thus in the cell proliferation, motility, invasion, and of human ovarian cancer cell line SKOV3.
Similar content being viewed by others
References
Chambers AF, Groom AC, MacDonald IC. Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer. 2002;2:563–72.
Hall A. Rho GTPases and the actin cytoskeleton. Science. 1998;279:509–14.
Mayer BJ, Hamaguchi M, Hanafusa H. A novel viral oncogene with structural similarity to phospholipase C.:. Nature. 1988;332(6161):272–5.
Matsuda M, Tanaka S, Nagata S, Kojima A, Kurata T, Shibuya M. Two species of human CRK cDNA encode proteins with distinct biological activities. Mol Cell Biol. 1992;12:3482–9.
Tanaka S, Hattori S, Kurata T, Nagashima K, Fukui Y, Nakamura S, et al. Both the SH2 and SH3 domains of human CRK protein are required for neuronal differentiation of PC12 cells. Mol Cell Biol. 1993;13:4409–15.
Gotoh T, Hattori S, Nakamura S, Kitayama H, Noda M, Takai Y, et al. Identification of Rap1 as a target for the Crk SH3 domain-binding guanine nucleotide-releasing factor C3G. Mol Cell Biol. 1995;15(12):6746–53.
Hasegawa H, Kiyokawa E, Tanaka S, Nagashima K, Gotoh N, Shibuya M, et al. DOCK180, a major CRK-binding protein, alters cell morphology upon translocation to the cell membrane. Mol Cell Biol. 1996;4:1770–6.
Kiyokawa E, Hashimoto Y, Kobayashi S, Sugimura H, Kurata T, Matsuda M. Activation of Rac1 by a Crk SH3-binding protein, DOCK180. Genes Dev. 1998;12:3331–6.
Nishihara H, Tanaka S, Tsuda M, Oikawa S, Maeda M, Shimizu M, et al. Molecular and immunohistochemical analysis of signaling adaptor protein Crk in human cancers. Cancer Lett. 2002;180(1):55–61.
Miller CT, Chen G, Gharib TG, Wang H, Thomas DG, Misek DE, et al. Increased C-CRK proto-oncogene expression is associated with an aggressive phenotype in lung adenocarcinomas. Oncogene. 2003;22(39):7950–7.
Takino T, Nakada M, Miyamori H, Yamashita J, Yamada KM, Sato H. CrkI adapter protein modulates cell migration and invasion in glioblastoma. Cancer Res. 2003;63(9):2335–7.
Linghu H, Tsuda M, Makino Y, Watanabe T, Ichihara S, Sawa H, et al. Involvement of adaptor protein Crk in malignant feature of human ovarian cancer cell line MCAS. Oncogene. 2006;25:3547–56.
Hasegawa H, Kiyokawa E, Tanaka S, et al. DOCK180, a major CRK-binding protein, alters cell morphology upon translocation to the cell membrane. Mol Cell Biol. 1996;16:1770–6.
Erickson MR, Galletta BJ, Abmayr SM. Drosophila myoblast city encodes a conserved protein that is essential for myoblast fusion, dorsal closure, and cytoskeletal organization. J Cell Biol. 1997;138:589–603.
Nolan KM, Barrett K, Lu Y, Hu K, Vincent S, Settleman J. Myoblast city, the Drosophila homolog of DOCK180/CED-5, is required in a Rac signaling pathway utilized for multiple developmental processes. Genes Dev. 1998;12:3337–42.
Wu YC, Horvitz HR. C. elegans phagocytosis and cellmigration protein CED-5 is similar to human DOCK180. Nature. 1998;392:501–4.
Gumienny TL, Brugnera E, Tosello-Trampont AC, Kinchen JM, Haney LB, Nishiwaki K. CED-12/ELMO, a novel member of the CrkII/Dock180/Rac pathway, is required for phagocytosis and cell migration. Cell. 2001;107(1):27–41.
Etienne-Manneville S, Hall A. Rho GTPases in cell biology. Nature. 2002;420:629–35.
Para A, Krischke M, Merlot S, Shen Z, Oberholzer M, Lee S, et al. Dictyostelium Dock180-related RacGEFs regulate the actin cytoskeleton during cell motility. Mol Biol Cell. 2009;20(2):699–707.
Lu M, Ravichandran KS. Dock180–ELMO cooperation in Rac activation. Methods Enzymol. 2006;406:388–402.
Grimsley CM, Kinchen JM, Tosello-Trampont A, Brugnera E, Haney LB, Lu M, et al. Dock180 and ELMO1 proteins cooperate to promote evolutionarily conserved Rac-dependent cell migration. J Biol Chem. 2004;279(7):6087–97.
Payne SL, Hendrix MJ, Kirschmann DA. Lysyl oxidase regulates actin filament formation through the p130(Cas)/Crk/DOCK180 signaling complex. J Cell Biochem. 2006;98(4):827–37.
Akakura S, Kar B, Singh S, Cho L, Tibrewal N, Sanokawa-Akakura R, et al. C-terminal SH3 domain of CrkII regulates the assembly and function of the DOCK180/ELMO Rac-GEF. J Cell Physiol. 2005;204(1):344–51.
Gumienny TL, Brugnera E, Tosello-Trampont AC, Kinchen JM, Haney LB, Nishiwaki K, et al. CED-12/ELMO, a novel member of the CrkII/Dock180/Rac pathway. Is required for phagocytosis and cell migration. Cell. 2001;107(1):27–41.
Brugnera E, Haney L, Grimsley C, Lu M, Walk SF, Tosello-Trampont AC, et al. Unconventional Rac-GEF activity is mediated through the Dock180–ELMO complex. Nat Cell Biol. 2002;4:574–82.
Albert ML, Kim JI, Birge RB. Alphavbeta5 integrin recruits the CrkII–Dock180–rac1 complex for phagocytosis of apoptotic cells. Nat Cell Biol. 2000;2:899–905.
Hsia DA, Mitra SK, Hauck CR, Streblow DN, Nelson JA, Ilic D, et al. Differential regulation of cell motility and invasion by FAK. Cell Biol. 2003;160(5):753–67.
Valle AM, Beuvin M, Boyer B. Activation of Rac1 by Paxillin–Crk–DOCK180 signaling complex is antagonized by Rap1 in migrating NBT-II cells. J Biol Chem. 2004;279(43):44940–6.
Gu J, Sumida Y, Sanzen N, Sekiguchi K. Laminin-10/11 and fibronectin differentially regulate integrin-dependent Rho and Rac activation via p130(Cas)–CrkII–DOCK180 pathway. J Biol Chem. 2001;276(29):27090–7.
Jarzynka MJ, Hu B, Hui KM, Bar-Joseph I, Gu W, Hirose T, et al. ELMO1 and Dock180, a bipartite Rac1 guanine nucleotide exchange factor, promote human glioma cell invasion. Cancer Res. 2007;67(15):7203–11.
Kiyokawa E, Hashimoto Y, Kurata T, Sugimura H, Matsuda M. Evidence that DOCK180 up-regulates signals from the CrkII–p130(Cas) complex. J Biol Chem. 1998;273(38):24479–84.
Tanaka S, Morishita T, Hashimoto Y, Hattori S, Nakamura S, Shibuya M, et al. C3G, a guanine nucleotide-releasing protein expressed ubiquitously, binds to the Src homology 3 domains of CRK and GRB2/ASH proteins. Proc Natl Acad Sci U S A. 1994;91(8):3443–7.
Kitayama H, Sugimoto Y, Matsuzaki T, Ikawa Y, Noda M. A ras-related gene with transformation suppressor activity. Cell. 1989;56(1):77–84.
Van Aelst L, Symons M. Role of Rho family GTPases in epithelial morphogenesis. Genes Dev. 2002;16(9):1032–54.
Bivona TG, Wiener HH, Ahearn IM, Silletti J, Chiu VK, Philips MR. Rap1 up-regulation and activation on plasma membrane regulates T cell adhesion. J Cell Biol. 2004;164(3):461–70.
Larue L, Bellacosa A. Epithelial-mesenchymal transition in development and cancer: role of phosphatidylinositol 3′ kinase/AKT pathways. Oncogene. 2005;24(50):7443–54.
Acknowledgments
We thank Dr. Qiou Wei in Center for Cancer Research, National Cancer Institute at Frederick, MD, USA, for his technical support. This work was supported by the grant from the National Natural Science Foundation of China (NSFC no. C30672432, no.C30772330) and was supported partly by the Natural Science Foundation Project of CQ CSTC, 2007BB5319, the grant for returned students abroad from the Ministry of Education of the People's Republic of China(2007), and the grant for Medical Sciences from the First Hospital, Chongqing Medical University (YXJJ2009-06).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary materials
ESM 1
(PDF 37 kb)
Rights and permissions
About this article
Cite this article
Wang, H., Linghu, H., Wang, J. et al. The role of Crk/Dock180/Rac1 pathway in the malignant behavior of human ovarian cancer cell SKOV3. Tumor Biol. 31, 59–67 (2010). https://doi.org/10.1007/s13277-009-0009-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13277-009-0009-9