title: WAVE2- and microtubule-dependent formation of long protrusions and invasion of cancer cells cultured on three-dimensional extracellular matrices.
abstract: Invadopodia, small protrusions formed at ventral membranes of several types of invasive cancer cells upon contact with the extracellular matrix (ECM), are implicated in cell invasion; however, the relationship between invadopodia formation and cell invasion through the ECM is still unknown. To correlate the formation of membrane protrusions and cell invasion, a three-dimensional (3-D) gel culture system with native collagen type-I matrix overlaid with a thin basement membrane equivalent (Matrigel) was made. Human breast cancer cell line MDA-MB-231 formed long protrusions in addition to small protrusions reminiscent of invadopodia and migrated into the collagen layer. Comparative analyses with other cancer cell lines indicate that cellular ability to form long protrusions, but not small protrusions or invadopodia, correlates with cellular invasiveness in the 3-D culture. Some of the long protrusions in MDA-MB-231 cells appeared to extend from the adherence membrane, implying that they are derived from small protrusions. The formation of long protrusions and invasion, as well as the formation of invadopodia, required WAVE2 in MDA-MB-231 cells. Accumulation of tubulin was observed in long protrusions but not in invadopodia. Correspondingly, a microtubule-stabilizing agent, paclitaxel, suppressed the formation of long protrusions and invasion, but not the formation of invadopodia, in MDA-MB-231 cells. These results suggest that long protrusions formed in a WAVE2- and microtubule-dependent manner may identify the cells at the later stage of invasion, possibly after the formation of invadopodia in the 3-D cultures.
authors.affiliation: Molecular Cell Biology Division, Kanagawa Cancer Center Research Institute, Yokohama, Japan. kei@gancen.asahi.yokohama.jp
authors.forename: Keiji
authors.initials: K
authors.lastname: Kikuchi
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D006801:Humans; D007797:Laminin / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology; D011509:Proteoglycans / Q000378:metabolism; D014407:Tumor Cells, Cultured; D051300:Wiskott-Aldrich Syndrome Protein Family / Q000378:metabolism*
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D051300:Wiskott-Aldrich Syndrome Protein Family; C059485:matrigel; D003094:Collagen
references.citation: Cancer Sci. 2007 Jul;98(7):992-9
references.pmid: 17459058
references.citation: Dev Cell. 2003 Oct;5(4):595-609
references.pmid: 14536061
references.citation: Eur J Cell Biol. 2006 Dec;85(12):1217-31
references.pmid: 17010475
references.citation: Nat Rev Mol Cell Biol. 2007 Jan;8(1):37-48
references.pmid: 17183359
references.citation: Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6647-52
references.pmid: 15087504
references.citation: Oncogene. 1999 Aug 5;18(31):4440-9
references.pmid: 10442635
references.citation: Curr Opin Cell Biol. 2007 Feb;19(1):88-94
references.pmid: 17194581
references.citation: Cancer Res. 1998 Aug 15;58(16):3743-50
references.pmid: 9721888
references.citation: Eur J Cell Biol. 2006 Apr;85(3-4):159-64
references.pmid: 16546558
references.citation: Cell. 2006 Nov 17;127(4):679-95
references.pmid: 17110329
references.citation: Oncogene. 2001 Oct 1;20(44):6418-34
references.pmid: 11607842
references.citation: Cancer Res. 2004 Dec 15;64(24):8924-31
references.pmid: 15604254
references.citation: Exp Cell Res. 2004 Aug 1;298(1):178-87
references.pmid: 15242772
references.citation: EMBO J. 2003 Jul 15;22(14):3602-12
references.pmid: 12853475
references.citation: Ann N Y Acad Sci. 1999 Jun 30;878:361-71
references.pmid: 10415741
references.citation: Nat Rev Mol Cell Biol. 2004 Aug;5(8):647-57
references.pmid: 15366708
references.citation: Nature. 1998 Jan 1;391(6662):93-6
references.pmid: 9422512
references.citation: Mol Biol Cell. 2000 Sep;11(9):2999-3012
references.pmid: 10982396
references.citation: Cancer Res. 2002 Feb 1;62(3):669-74
references.pmid: 11830518
references.citation: Nat Rev Cancer. 2003 May;3(5):362-74
references.pmid: 12724734
references.citation: Clin Exp Metastasis. 2006;23(2):97-105
references.pmid: 16830222
references.citation: Clin Cancer Res. 2006 Oct 1;12(19):5673-9
references.pmid: 17020969
references.citation: J Cell Biol. 2003 Jan 20;160(2):267-77
references.pmid: 12527751
references.citation: Oncogene. 2005 Feb 17;24(8):1309-19
references.pmid: 15608687
references.citation: Exp Cell Res. 2008 Jul 1;314(11-12):2313-22
references.pmid: 18514191
references.citation: Exp Cell Res. 2006 May 1;312(8):1240-53
references.pmid: 16442522
references.citation: Curr Biol. 2004 Apr 20;14(8):697-703
references.pmid: 15084285
references.citation: Front Biosci. 2004 May 01;9:1944-53
references.pmid: 14977600
references.citation: Cancer Res. 2006 Mar 15;66(6):3034-43
references.pmid: 16540652
references.citation: J Cell Biol. 2005 Jan 31;168(3):441-52
references.pmid: 15684033
references.citation: Nature. 2000 Dec 7;408(6813):732-5
references.pmid: 11130076
references.citation: Nat Rev Mol Cell Biol. 2002 Dec;3(12):957-64
references.pmid: 12461561
references.citation: J Exp Zool. 1989 Aug;251(2):167-85
references.pmid: 2549171
references.citation: Semin Cancer Biol. 2008 Feb;18(1):23-34
references.pmid: 17936638
references.citation: Nat Cell Biol. 1999 May;1(1):45-50
references.pmid: 10559863
references.citation: Curr Opin Cell Biol. 2005 Oct;17(5):524-32
references.pmid: 16112853
references.citation: Cancer Res. 2007 May 1;67(9):4227-35
references.pmid: 17483334
references.citation: Clin Cancer Res. 1998 Feb;4(2):507-15
references.pmid: 9516943
references.citation: Cells Tissues Organs. 2000;167(1):1-8
references.pmid: 10899710
references.citation: Nature. 2003 Jul 24;424(6947):452-6
references.pmid: 12879075
references.citation: EMBO J. 2005 Mar 9;24(5):963-73
references.pmid: 15719014
references.citation: Methods Enzymol. 2006;406:625-43
references.pmid: 16472693