Actin bundling by dynamin 2 and cortactin is implicated in cell migration by stabilizing filopodia in human non-small cell lung carcinoma cells

Yamada,Hiroshi; Takeda,Tetsuya; Michiue,Hiroyuki; Abe,Tadashi; Takei,Kohji

doi:10.3892/ijo.2016.3592

Actin bundling by dynamin 2 and cortactin is implicated in cell migration by stabilizing filopodia in human non-small cell lung carcinoma cells

Authors:
- Hiroshi Yamada
- Tetsuya Takeda
- Hiroyuki Michiue
- Tadashi Abe
- Kohji Takei
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Affiliations: Department of Neuroscience, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Kita-ku, Okayama 700-8558, Japan, Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Kita-ku, Okayama 700-8558, Japan
Published online on: June 30, 2016 https://doi.org/10.3892/ijo.2016.3592
Pages: 877-886
Copyright: © Yamada et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The endocytic protein dynamin participates in the formation of actin-based membrane protrusions such as podosomes, pseudopodia, and invadopodia, which facilitate cancer cell migration, invasion, and metastasis. However, the role of dynamin in the formation of actin-based membrane protrusions at the leading edge of cancer cells is unclear. In this study, we demonstrate that the ubiquitously expressed dynamin 2 isoform facilitates cell migration by stabilizing F-actin bundles in filopodia of the lung cancer cell line H1299. Pharmacological inhibition of dynamin 2 decreased cell migration and filopodial formation. Furthermore, dynamin 2 and cortactin mostly colocalized along F-actin bundles in filopodia of serum-stimulated H1299 cells by immunofluorescent and immunoelectron microscopy. Knockdown of dynamin 2 or cortactin inhibited the formation of filopodia in serum-stimulated H1299 cells, concomitant with a loss of F-actin bundles. Expression of wild-type cortactin rescued the punctate-like localization of dynamin 2 and filopodial formation. The incubation of dynamin 2 and cortactin with F-actin induced the formation of long and thick actin bundles, with these proteins colocalizing at F-actin bundles. A depolymerization assay revealed that dynamin 2 and cortactin increased the stability of F-actin bundles. These results indicate that dynamin 2 and cortactin participate in cell migration by stabilizing F-actin bundles in filopodia. Taken together, these findings suggest that dynamin might be a possible molecular target for anticancer therapy.

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1	Arjonen A, Kaukonen R and Ivaska J: Filopodia and adhesion in cancer cell motility. Cell Adhes Migr. 5:421–430. 2011. View Article : Google Scholar
2	Ridley AJ: Life at the leading edge. Cell. 145:1012–1022. 2011. View Article : Google Scholar : PubMed/NCBI
3	Takei K, Slepnev VI, Haucke V and De Camilli P: Functional partnership between amphiphysin and dynamin in clathrin-mediated endocytosis. Nat Cell Biol. 1:33–39. 1999.PubMed/NCBI
4	Mettlen M, Pucadyil T, Ramachandran R and Schmid SL: Dissecting dynamin’s role in clathrin-mediated endocytosis. Biochem Soc Trans. 37:1022–1026. 2009. View Article : Google Scholar : PubMed/NCBI
5	Praefcke GJ and McMahon HT: The dynamin superfamily: Universal membrane tubulation and fission molecules? Nat Rev Mol Cell Biol. 5:133–147. 2004. View Article : Google Scholar : PubMed/NCBI
6	Cao H, Garcia F and McNiven MA: Differential distribution of dynamin isoforms in mammalian cells. Mol Biol Cell. 9:2595–2609. 1998. View Article : Google Scholar : PubMed/NCBI
7	McNiven MA, Kim L, Krueger EW, Orth JD, Cao H and Wong TW: Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. J Cell Biol. 151:187–198. 2000. View Article : Google Scholar : PubMed/NCBI
8	Baldassarre M, Pompeo A, Beznoussenko G, Castaldi C, Cortellino S, McNiven MA, Luini A and Buccione R: Dynamin participates in focal extracellular matrix degradation by invasive cells. Mol Biol Cell. 14:1074–1084. 2003. View Article : Google Scholar : PubMed/NCBI
9	Ochoa GC, Slepnev VI, Neff L, Ringstad N, Takei K, Daniell L, Kim W, Cao H, McNiven M, Baron R, et al: A functional link between dynamin and the actin cytoskeleton at podosomes. J Cell Biol. 150:377–389. 2000. View Article : Google Scholar : PubMed/NCBI
10	Torre E, McNiven MA and Urrutia R: Dynamin 1 antisense oligonucleotide treatment prevents neurite formation in cultured hippocampal neurons. J Biol Chem. 269:32411–32417. 1994.PubMed/NCBI
11	Kurklinsky S, Chen J and McNiven MA: Growth cone morphology and spreading are regulated by a dynamin-cortactin complex at point contacts in hippocampal neurons. J Neurochem. 117:48–60. 2011. View Article : Google Scholar : PubMed/NCBI
12	Yamada H, Abe T, Satoh A, Okazaki N, Tago S, Kobayashi K, Yoshida Y, Oda Y, Watanabe M, Tomizawa K, et al: Stabilization of actin bundles by a dynamin 1/cortactin ring complex is necessary for growth cone filopodia. J Neurosci. 33:4514–4526. 2013. View Article : Google Scholar : PubMed/NCBI
13	Gold ES, Underhill DM, Morrissette NS, Guo J, McNiven MA and Aderem A: Dynamin 2 is required for phagocytosis in macrophages. J Exp Med. 190:1849–1856. 1999. View Article : Google Scholar : PubMed/NCBI
14	Otsuka A, Abe T, Watanabe M, Yagisawa H, Takei K and Yamada H: Dynamin 2 is required for actin assembly in phagocytosis in Sertoli cells. Biochem Biophys Res Commun. 378:478–482. 2009. View Article : Google Scholar
15	Faelber K, Posor Y, Gao S, Held M, Roske Y, Schulze D, Haucke V, Noé F and Daumke O: Crystal structure of nucleotide-free dynamin. Nature. 477:556–560. 2011. View Article : Google Scholar : PubMed/NCBI
16	Ford MG, Jenni S and Nunnari J: The crystal structure of dynamin. Nature. 477:561–566. 2011. View Article : Google Scholar : PubMed/NCBI
17	Wu H, Reynolds AB, Kanner SB, Vines RR and Parsons JT: Identification and characterization of a novel cytoskeleton-associated pp60src substrate. Mol Cell Biol. 11:5113–5124. 1991. View Article : Google Scholar : PubMed/NCBI
18	MacGrath SM and Koleske AJ: Cortactin in cell migration and cancer at a glance. J Cell Sci. 125:1621–1626. 2012. View Article : Google Scholar : PubMed/NCBI
19	Ammer AG and Weed SA: Cortactin branches out: Roles in regulating protrusive actin dynamics. Cell Motil Cytoskeleton. 65:687–707. 2008. View Article : Google Scholar : PubMed/NCBI
20	Yamada H, Abe T, Li SA, Masuoka Y, Isoda M, Watanabe M, Nasu Y, Kumon H, Asai A and Takei K: Dynasore, a dynamin inhibitor, suppresses lamellipodia formation and cancer cell invasion by destabilizing actin filaments. Biochem Biophys Res Commun. 390:1142–1148. 2009. View Article : Google Scholar : PubMed/NCBI
21	Yamada H, Abe T, Li SA, Tago S, Huang P, Watanabe M, Ikeda S, Ogo N, Asai A and Takei K: N′-[4-(dipropylamino)benzylidene]-2-hydroxybenzohydrazide is a dynamin GTPase inhibitor that suppresses cancer cell migration and invasion by inhibiting actin polymerization. Biochem Biophys Res Commun. 443:511–517. 2014. View Article : Google Scholar
22	Mooren OL, Kotova TI, Moore AJ and Schafer DA: Dynamin2 GTPase and cortactin remodel actin filaments. J Biol Chem. 284:23995–24005. 2009. View Article : Google Scholar : PubMed/NCBI
23	Masaike Y, Takagi T, Hirota M, Yamada J, Ishihara S, Yung TM, Inoue T, Sawa C, Sagara H, Sakamoto S, et al: Identification of dynamin-2-mediated endocytosis as a new target of osteoporosis drugs, bisphosphonates. Mol Pharmacol. 77:262–269. 2010. View Article : Google Scholar
24	Slepnev VI, Ochoa GC, Butler MH and De Camilli P: Tandem arrangement of the clathrin and AP-2 binding domains in amphiphysin 1 and disruption of clathrin coat function by amphiphysin fragments comprising these sites. J Biol Chem. 275:17583–17589. 2000. View Article : Google Scholar : PubMed/NCBI
25	Eppinga RD, Krueger EW, Weller SG, Zhang L, Cao H and McNiven MA: Increased expression of the large GTPase dynamin 2 potentiates metastatic migration and invasion of pancreatic ductal carcinoma. Oncogene. 31:1228–1241. 2012. View Article : Google Scholar
26	Macia E, Ehrlich M, Massol R, Boucrot E, Brunner C and Kirchhausen T: Dynasore, a cell-permeable inhibitor of dynamin. Dev Cell. 10:839–850. 2006. View Article : Google Scholar : PubMed/NCBI
27	Hill TA, Gordon CP, McGeachie AB, Venn-Brown B, Odell LR, Chau N, Quan A, Mariana A, Sakoff JA, Chircop M, et al: Inhibition of dynamin mediated endocytosis by the dynoles--synthesis and functional activity of a family of indoles. J Med Chem. 52:3762–3773. 2009. View Article : Google Scholar : PubMed/NCBI
28	Quan A, McGeachie AB, Keating DJ, van Dam EM, Rusak J, Chau N, Malladi CS, Chen C, McCluskey A, Cousin MA, et al: Myristyl trimethyl ammonium bromide and octadecyl trimethyl ammonium bromide are surface-active small molecule dynamin inhibitors that block endocytosis mediated by dynamin I or dynamin II. Mol Pharmacol. 72:1425–1439. 2007. View Article : Google Scholar : PubMed/NCBI
29	Schafer DA, Weed SA, Binns D, Karginov AV, Parsons JT and Cooper JA: Dynamin 2 and cortactin regulate actin assembly and filament organization. Curr Biol. 12:1852–1857. 2002. View Article : Google Scholar : PubMed/NCBI
30	Takei K, McPherson PS, Schmid SL and De Camilli P: Tubular membrane invaginations coated by dynamin rings are induced by GTP-gamma S in nerve terminals. Nature. 374:186–190. 1995. View Article : Google Scholar : PubMed/NCBI
31	Sweitzer SM and Hinshaw JE: Dynamin undergoes a GTP-dependent conformational change causing vesiculation. Cell. 93:1021–1029. 1998. View Article : Google Scholar : PubMed/NCBI
32	Takei K, Haucke V, Slepnev V, Farsad K, Salazar M, Chen H and De Camilli P: Generation of coated intermediates of clathrin-mediated endocytosis on protein-free liposomes. Cell. 94:131–141. 1998. View Article : Google Scholar : PubMed/NCBI
33	Roux A, Uyhazi K, Frost A and De Camilli P: GTP-dependent twisting of dynamin implicates constriction and tension in membrane fission. Nature. 441:528–531. 2006. View Article : Google Scholar : PubMed/NCBI
34	Feng H, Liu KW, Guo P, Zhang P, Cheng T, McNiven MA, Johnson GR, Hu B and Cheng SY: Dynamin 2 mediates PDGFRα-SHP-2-promoted glioblastoma growth and invasion. Oncogene. 31:2691–2702. 2012. View Article : Google Scholar