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BMC Cancer
Published in: BMC Cancer 1/2013

Open Access 01-12-2013 | Research article

APC/β-catenin-rich complexes at membrane protrusions regulate mammary tumor cell migration and mesenchymal morphology

Authors: Matthew A Odenwald, Jenifer R Prosperi, Kathleen H Goss

Published in: BMC Cancer | Issue 1/2013

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Abstract

Background

The APC tumor suppressor is mutated or downregulated in many tumor types, and is prominently localized to punctate clusters at protrusion tips in migratory cells, such as in astrocytes where it has been implicated in directed cell motility. Although APC loss is considered an initiating event in colorectal cancer, for example, it is less clear what role APC plays in tumor cell motility and whether loss of APC might be an important promoter of tumor progression in addition to initiation.

Methods

The localization of APC and β-catenin was analyzed in multiple cell lines, including non-transformed epithelial lines treated with a proteasome inhibitor or TGFβ to induce an epithelial-to-mesenchymal transition (EMT), as well as several breast cancer lines, by immunofluorescence. APC expression was knocked down in 4T07 mammary tumor cells using lentiviral-mediated delivery of APC-specific short-hairpin (sh) RNAs, and assessed using quantitative (q) reverse-transcriptase (RT)-PCR and western blotting. Tumor cell motility was analyzed by performing wound-filling assays, and morphology via immunofluorescence (IF) and phase-contrast microscopy. Additionally, proliferation was measured using BrdU incorporation, and TCF reporter assays were performed to determine β-catenin/TCF-mediated transcriptional activity.

Results

APC/β-catenin-rich complexes were observed at protrusion ends of migratory epithelial cells treated with a proteasome inhibitor or when EMT has been induced and in tumor cells with a mesenchymal, spindle-like morphology. 4T07 tumor cells with reduced APC levels were significantly less motile and had a more rounded morphology; yet, they did not differ significantly in proliferation or β-catenin/TCF transcriptional activity. Furthermore, we found that APC/β-catenin-rich complexes at protrusion ends were dependent upon an intact microtubule cytoskeleton.

Conclusions

These findings indicate that membrane protrusions with APC/β-catenin-containing puncta control the migratory potential and mesenchymal morphology of mammary tumor cells and suggest that APC loss during later stages of tumor progression might impact tumor cell dissemination or colonization.
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Literature
1.
Groden J, Thliveris A, Samowitz W, Carlson M, Gelbert L, Albertsen H, Joslyn G, Stevens J, Spirio L, Robertson M, et al: Identification and characterization of the familial adenomatous polyposis coli gene. Cell. 1991, 66: 589-600. 10.1016/0092-8674(81)90021-0.CrossRefPubMed
2.
Kinzler K, Nilbert MC, Su L-K, Vogelstein B, Bryan TM, Levy DB, Smith KJ, Preisinger AM, Hedge P, McKechnie D, et al: Identification of FAP locus genes from chromosome 5q21. Science. 1991, 253: 661-665. 10.1126/science.1651562.CrossRefPubMed
3.
Nishisho I, Nakamura Y, Miyoshi Y, Miki Y, Ando H, Horii A, Koyama K, Utsunomiya J, Baba S, Hedge P, et al: Mutations of chromosome 5q21 genes in FAP and colorectal cancer patients. Science. 1991, 253: 665-669. 10.1126/science.1651563.CrossRefPubMed
4.
Prosperi JR, Lue HH, Goss KH: Dysregulation of the WNT Pathway in Solid Tumors. Targeting the Wnt Pathway in Cancer. Edited by: Goss K, Kahn M. 2011, New York: Springer, 81-128.CrossRef
5.
Prosperi JR, Becher KR, Willson TA, Collins MH, Witte DP, Goss KH: The APC tumor suppressor is required for epithelial integrity in the mouse mammary gland. J Cell Physiol. 2009, 220 (2): 319-331. 10.1002/jcp.21766.CrossRefPubMed
6.
Prosperi JR, Khramtsov AI, Khramtsova GF, Goss KH: Apc mutation enhances PyMT-induced mammary tumorigenesis. PLoS One. 2011, 6 (12): e29339-10.1371/journal.pone.0029339.CrossRefPubMedPubMedCentral
7.
Vlad A, Rohrs S, Klein-Hitpass L, Muller O: The first five years of the Wnt targetome. Cell Signal. 2008, 20 (5): 795-802. 10.1016/j.cellsig.2007.10.031.CrossRefPubMed
8.
Nathke IS: The adenomatous polyposis coli protein: the Achilles heel of the gut epithelium. Annu Rev Cell Dev Biol. 2004, 20: 337-366. 10.1146/annurev.cellbio.20.012103.094541.CrossRefPubMed
9.
Prosperi JR, Goss KH: Wnt Pathway-Independent Activities of the APC Tumor Suppressor. Tumor Suppressors. Edited by: Nguyen SD. 2011, Hauppauge: Nova Science Publishers
10.
Nathke IS, Adams CL, Polakis P, Sellin JH, Nelson WJ: The adenomatous polyposis coli tumor suppressor protein localizes to plasma membrane sites involved in active cell migration. J Cell Biol. 1996, 134 (1): 165-179. 10.1083/jcb.134.1.165.CrossRefPubMed
11.
Kroboth K, Newton IP, Kita K, Dikovskaya D, Zumbrunn J, Waterman-Storer CM, Nathke IS: Lack of adenomatous polyposis coli protein correlates with a decrease in cell migration and overall changes in microtubule stability. Mol Biol Cell. 2007, 18 (3): 910-918. 10.1091/mbc.E06-03-0179.CrossRefPubMedPubMedCentral
12.
Iizuka-Kogo A, Shimomura A, Senda T: Colocalization of APC and DLG at the tips of cellular protrusions in cultured epithelial cells and its dependency on cytoskeletons. Histochem Cell Biol. 2005, 123 (1): 67-73. 10.1007/s00418-004-0729-2.CrossRefPubMed
13.
Watanabe T, Wang S, Noritake J, Sato K, Fukata M, Takefuji M, Nakagawa M, Izumi N, Akiyama T, Kaibuchi K: Interaction with IQGAP1 links APC to Rac1, Cdc42, and actin filaments during cell polarization and migration. Dev Cell. 2004, 7 (6): 871-883. 10.1016/j.devcel.2004.10.017.CrossRefPubMed
14.
Etienne-Manneville S, Hall A: Cdc42 regulates GSK-3beta and adenomatous polyposis coli to control cell polarity. Nature. 2003, 421 (6924): 753-756. 10.1038/nature01423.CrossRefPubMed
15.
Etienne-Manneville S, Manneville JB, Nicholls S, Ferenczi MA, Hall A: Cdc42 and Par6-PKCzeta regulate the spatially localized association of Dlg1 and APC to control cell polarization. J Cell Biol. 2005, 170 (6): 895-901. 10.1083/jcb.200412172.CrossRefPubMedPubMedCentral
16.
Fialka I, Pasquali C, Lottspeich F, Ahorn H, Huber LA: Subcellular fractionation of polarized epithelial cells and identification of organelle-specific proteins by two-dimensional gel electrophoresis. Electrophoresis. 1997, 18 (14): 2582-2590. 10.1002/elps.1150181414.CrossRefPubMed
17.
Matlin KS, Reggio H, Helenius A, Simons K: Infectious entry pathway of influenza virus in a canine kidney cell line. J Cell Biol. 1981, 91 (3 Pt 1): 601-613.CrossRefPubMed
18.
Aslakson CJ, Miller FR: Selective events in the metastatic process defined by analysis of the sequential dissemination of subpopulations of a mouse mammary tumor. Cancer Res. 1992, 52 (6): 1399-1405.PubMed
19.
Wang Y, Azuma Y, Friedman DB, Coffey RJ, Neufeld KL: Novel association of APC with intermediate filaments identified using a new versatile APC antibody. BMC Cell Biol. 2009, 10: 75-10.1186/1471-2121-10-75.CrossRefPubMedPubMedCentral
20.
Barth AI, Pollack AL, Altschuler Y, Mostov KE, Nelson WJ: NH2-terminal deletion of beta-catenin results in stable colocalization of mutant beta-catenin with adenomatous polyposis coli protein and altered MDCK cell adhesion. J Cell Biol. 1997, 136 (3): 693-706. 10.1083/jcb.136.3.693.CrossRefPubMedPubMedCentral
21.
Kita K, Wittmann T, Nathke IS, Waterman-Storer CM: Adenomatous polyposis coli on microtubule plus ends in cell extensions can promote microtubule net growth with or without EB1. Mol Biol Cell. 2006, 17 (5): 2331-2345. 10.1091/mbc.E05-06-0498.CrossRefPubMedPubMedCentral
22.
Heinen CD, Goss KH, Cornelius JR, Babcock GF, Knudsen ES, Kowalik T, Groden J: The APC tumor suppressor controls entry into S-phase through its ability to regulate the cyclin D/RB pathway. Gastroenterology. 2002, 123: 751-763. 10.1053/gast.2002.35382.CrossRefPubMed
23.
Mogensen MM, Tucker JB, Mackie JB, Prescott AR, Nathke IS: The adenomatous polyposis coli protein unambiguously localizes to microtubule plus ends and is involved in establishing parallel arrays of microtubule bundles in highly polarized epithelial cells. J Cell Biol. 2002, 157 (6): 1041-1048. 10.1083/jcb.200203001.CrossRefPubMedPubMedCentral
24.
Berrueta L, Kraeft SK, Tirnauer JS, Schuyler SC, Chen LB, Hill DE, Pellman D, Bierer BE: The adenomatous polyposis coli-binding protein EB1 is associated with cytoplasmic and spindle microtubules. Proc Natl Acad Sci USA. 1998, 95 (18): 10596-10601. 10.1073/pnas.95.18.10596.CrossRefPubMedPubMedCentral
25.
Mimori-Kiyosue Y, Shiina N, Tsukita S: Adenomatous polyposis coli (APC) protein moves along microtubules and concentrates at their growing ends in epithelial cells. J Cell Biol. 2000, 148 (3): 505-518. 10.1083/jcb.148.3.505.CrossRefPubMedPubMedCentral
26.
Morrison EE, Wardleworth BN, Askham JM, Markham AF, Meredith DM: EB1, a protein which interacts with the APC tumour suppressor, is associated with the microtubule cytoskeleton throughout the cell cycle. Oncogene. 1998, 17 (26): 3471-3477.CrossRefPubMed
27.
Wen Y, Eng CH, Schmoranzer J, Cabrera-Poch N, Morris EJ, Chen M, Wallar BJ, Alberts AS, Gundersen GG: EB1 and APC bind to mDia to stabilize microtubules downstream of Rho and promote cell migration. Nat Cell Biol. 2004, 6 (9): 820-830. 10.1038/ncb1160.CrossRefPubMed
28.
Harris ES, Nelson WJ: Adenomatous polyposis coli regulates endothelial cell migration independent of roles in beta-catenin signaling and cell-cell adhesion. Mol Biol Cell. 2010, 21 (15): 2611-2623. 10.1091/mbc.E10-03-0235.CrossRefPubMedPubMedCentral
29.
Shi SH, Cheng T, Jan LY, Jan YN: APC and GSK-3beta are involved in mPar3 targeting to the nascent axon and establishment of neuronal polarity. Curr Biol: CB. 2004, 14 (22): 2025-2032. 10.1016/j.cub.2004.11.009.CrossRefPubMed
30.
Jimbo T, Kawasaki Y, Koyama R, Sato R, Takada S, Haraguchi K, Akiyama T: Identification of a link between the tumour suppressor APC and the kinesin superfamily. Nat Cell Biol. 2002, 4 (4): 323-327. 10.1038/ncb779.CrossRefPubMed
31.
Munemitsu S, Souza B, Muller O, Albert I, Rubinfeld B, Polakis P: The APC gene product associates with microtubules in vivo and promotes their assembly in vitro. Cancer Res. 1994, 54 (14): 3676-3681.PubMed
32.
Su LK, Burrell M, Hill DE, Gyuris J, Brent R, Wiltshire R, Trent J, Vogelstein B, Kinzler KW: APC binds to the novel protein EB1. Cancer Res. 1995, 55 (14): 2972-2977.PubMed
33.
Mili S, Moissoglu K, Macara IG: Genome-wide screen reveals APC-associated RNAs enriched in cell protrusions. Nature. 2008, 453 (7191): 115-119. 10.1038/nature06888.CrossRefPubMedPubMedCentral
34.
Mahmoud NN, Boolbol SK, Bilinski RT, Martucci C, Chadburn A, Bertagnolli MM: Apc gene mutation is associated with a dominant-negative effect upon intestinal cell migration. Cancer Res. 1997, 57 (22): 5045-5050.PubMed
35.
Sansom OJ, Reed KR, Hayes AJ, Ireland H, Brinkmann H, Newton IP, Batlle E, Simon-Assmann P, Clevers H, Nathke IS, et al: Loss of Apc in vivo immediately perturbs Wnt signaling, differentiation, and migration. Genes Dev. 2004, 18 (12): 1385-1390. 10.1101/gad.287404.CrossRefPubMedPubMedCentral
36.
Wong MH, Hermiston ML, Syder AJ, Gordon JI: Forced expression of the tumor suppressor adenomatosis polyposis coli protein induces disordered cell migration in the intestinal epithelium. Proc Natl Acad Sci USA. 1996, 93 (18): 9588-9593. 10.1073/pnas.93.18.9588.CrossRefPubMedPubMedCentral
37.
Faux MC, Ross JL, Meeker C, Johns T, Ji H, Simpson RJ, Layton MJ, Burgess AW: Restoration of full-length adenomatous polyposis coli (APC) protein in a colon cancer cell line enhances cell adhesion. J Cell Sci. 2004, 117 (Pt 3): 427-439.PubMed
38.
Kim K, Lu Z, Hay ED: Direct evidence for a role of beta-catenin/LEF-1 signaling pathway in induction of EMT. Cell Biol Int. 2002, 26 (5): 463-476. 10.1006/cbir.2002.0901.CrossRefPubMed
39.
Liebner S, Cattelino A, Gallini R, Rudini N, Iurlaro M, Piccolo S, Dejana E: Beta-catenin is required for endothelial-mesenchymal transformation during heart cushion development in the mouse. J Cell Biol. 2004, 166 (3): 359-367. 10.1083/jcb.200403050.CrossRefPubMedPubMedCentral
40.
Muller T, Bain G, Wang X, Papkoff J: Regulation of epithelial cell migration and tumor formation by beta-catenin signaling. Exp Cell Res. 2002, 280 (1): 119-133. 10.1006/excr.2002.5630.CrossRefPubMed
41.
Vincan E, Barker N: The upstream components of the Wnt signalling pathway in the dynamic EMT and MET associated with colorectal cancer progression. Clin Exp Metastasis. 2008, 25 (6): 657-663. 10.1007/s10585-008-9156-4.CrossRefPubMed
42.
Moseley JB, Bartolini F, Okada K, Wen Y, Gundersen GG, Goode BL: Regulated binding of adenomatous polyposis coli protein to actin. J Biol Chem. 2007, 282 (17): 12661-12668. 10.1074/jbc.M610615200.CrossRefPubMed
43.
Okada K, Bartolini F, Deaconescu AM, Moseley JB, Dogic Z, Grigorieff N, Gundersen GG, Goode BL: Adenomatous polyposis coli protein nucleates actin assembly and synergizes with the formin mDia1. J Cell Biol. 2010, 189 (7): 1087-1096. 10.1083/jcb.201001016.CrossRefPubMedPubMedCentral
44.
Kawasaki Y, Sagara M, Shibata Y, Shirouzu M, Yokoyama S, Akiyama T: Identification and characterization of Asef2, a guanine-nucleotide exchange factor specific for Rac1 and Cdc42. Oncogene. 2007, 26 (55): 7620-7267. 10.1038/sj.onc.1210574.CrossRefPubMed
45.
Kawasaki Y, Sato R, Akiyama T: Mutated APC and Asef are involved in the migration of colorectal tumour cells. Nat Cell Biol. 2003, 5 (3): 211-215. 10.1038/ncb937.CrossRefPubMed
46.
Kawasaki Y, Senda T, Ishidate T, Koyama R, Morishita T, Iwayama Y, Higuchi O, Akiyama T: Asef, a link between the tumor suppressor APC and G-protein signaling. Science. 2000, 289 (5482): 1194-1197. 10.1126/science.289.5482.1194.CrossRefPubMed
47.
Ashton GH, Morton JP, Myant K, Phesse TJ, Ridgway RA, Marsh V, Wilkins JA, Athineos D, Muncan V, Kemp R, et al: Focal adhesion kinase is required for intestinal regeneration and tumorigenesis downstream of Wnt/c-Myc signaling. Dev Cell. 2010, 19 (2): 259-269. 10.1016/j.devcel.2010.07.015.CrossRefPubMedPubMedCentral
48.
Faux MC, Coates JL, Kershaw NJ, Layton MJ, Burgess AW: Independent interactions of phosphorylated beta-catenin with E-cadherin at cell-cell contacts and APC at cell protrusions. PLoS One. 2010, 5 (11): e14127-10.1371/journal.pone.0014127.CrossRefPubMedPubMedCentral
49.
Sadot E, Conacci-Sorrell M, Zhurinsky J, Shnizer D, Lando Z, Zharhary D, Kam Z, Ben-Ze'ev A, Geiger B: Regulation of S33/S37 phosphorylated beta-catenin in normal and transformed cells. J Cell Sci. 2002, 115 (Pt 13): 2771-2780.PubMed
50.
Osmani N, Vitale N, Borg JP, Etienne-Manneville S: Scrib controls Cdc42 localization and activity to promote cell polarization during astrocyte migration. Curr Biol: CB. 2006, 16 (24): 2395-2405. 10.1016/j.cub.2006.10.026.CrossRefPubMed
51.
Heppner GH, Miller FR, Shekhar PM: Nontransgenic models of breast cancer. Breast Cancer Res. 2000, 2 (5): 331-334. 10.1186/bcr77.CrossRefPubMed
52.
Prasad CP, Mirza S, Sharma G, Prashad R, DattaGupta S, Rath G, Ralhan R: Epigenetic alterations of CDH1 and APC genes: relationship with activation of Wnt/beta-catenin pathway in invasive ductal carcinoma of breast. Life Sci. 2008, 83 (9–10): 318-325.CrossRefPubMed
53.
Hsieh JS, Lin SR, Chang MY, Chen FM, Lu CY, Huang TJ, Huang YS, Huang CJ, Wang JY: APC, K-ras, and p53 gene mutations in colorectal cancer patients: correlation to clinicopathologic features and postoperative surveillance. Am Surg. 2005, 71 (4): 336-343.PubMed
54.
Pecina-Slaus N, Majic Z, Musani V, Zeljko M, Cupic H: Report on mutation in exon 15 of the APC gene in a case of brain metastasis. J Neurooncol. 2010, 97 (1): 143-148. 10.1007/s11060-009-0001-7.CrossRefPubMed
55.
Pecina-Slaus N, Nikuseva Martic T, Zeljko M, Bulat S: Brain metastases exhibit gross deletions of the APC gene. Brain Tumor Pathol. 2011, 28 (3): 223-228. 10.1007/s10014-011-0030-8.CrossRefPubMed
Metadata
Title
APC/β-catenin-rich complexes at membrane protrusions regulate mammary tumor cell migration and mesenchymal morphology
Authors
Matthew A Odenwald
Jenifer R Prosperi
Kathleen H Goss
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2013
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-13-12

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