The online version of this article (doi:10.1186/1476-4598-11-36) contains supplementary material, which is available to authorized users.
The authors declare they have no competing interests.
DAT designed and performed experiments, analysed the data, and wrote the manuscript. JT performed the cell titre glo cell viability assay. JDB conceived of the study and wrote the manuscript. All authors read and approved the final manuscript.
The extracellular matrix (ECM) has a key role in facilitating the progression of ovarian cancer and we have shown recently that the secreted ECM protein TGFBI modulates the response of ovarian cancer to paclitaxel-induced cell death.
We have determined TGFBI signaling from the extracellular environment is preferential for the cell surface αvβ3 integrin heterodimer, in contrast to periostin, a TGFBI paralogue, which signals primarily via a β1 integrin-mediated pathway. We demonstrate that suppression of β1 integrin expression, in β3 integrin-expressing ovarian cancer cells, increases adhesion to rTGFBI. In addition, Syndecan-1 and −4 expression is dispensable for adhesion to rTGFBI and loss of Syndecan-1 cooperates with the loss of β1 integrin to further enhance adhesion to rTGFBI. The RGD motif present in the carboxy-terminus of TGFBI is necessary, but not sufficient, for SKOV3 cell adhesion and is dispensable for adhesion of ovarian cancer cells lacking β3 integrin expression. In contrast to TGFBI, the carboxy-terminus of periostin, lacking a RGD motif, is unable to support adhesion of ovarian cancer cells. Suppression of β3 integrin in SKOV3 cells increases resistance to paclitaxel-induced cell death while suppression of β1 integrin has no effect. Furthermore, suppression of TGFBI expression stimulates a paclitaxel resistant phenotype while suppression of fibronectin expression, which primarily signals through a β1 integrin-mediated pathway, increases paclitaxel sensitivity.
Therefore, different ECM components use distinct signaling mechanisms in ovarian cancer cells and in particular, TGFBI preferentially interacts through a β3 integrin receptor mediated mechanism to regulate the response of cells to paclitaxel-induced cell death.
Additional file 1: Movie S1. Bright-field time lapse video microscopy of an SKOV3 cell plated on rTGFBI in serum-free media. Images were acquired every 2 minutes for a period of 6 hours. (TIFF )12943_2011_1019_MOESM1_ESM.avi
Additional file 2: Movie S2. Bright-field time lapse video microscopy of an SKOV3 cell plated on fibronectin in serum-free media. Images were acquired every 2 minutes for a period of 6 hours. (TIFF )12943_2011_1019_MOESM2_ESM.avi
Additional file 3: Figure S1. Flow cytometric analysis of SKOV3, PEO1, and TR125 cell surface integrin expression following immunostaining with either IgG control or integrin-specific antibodies (P5D2 - β1, LM609 - αvβ3, P1F6 - αvβ5). (TIFF 700 KB)12943_2011_1019_MOESM3_ESM.tiff
Additional file 4: Figure S2. PEO1 cell adhesion to fibronectin, rTGFBI, rPOSTN and vitronectin coated tissue culture plastic in the presence of vehicle or the indicated integrin receptor blocking antibodies. Results are represented as the mean of two independent experiments normalized to uncoated and poly-L-lysine coated wells and represented as percent of vehicle treated control. Error bars represent the standard deviation. (TIFF 629 KB)12943_2011_1019_MOESM4_ESM.tiff
Additional file 5: Figure S3. SKOV3 cells expressing either control shRNA or β1 shRNA were left untreated or pretreated with the αvβ3 blocking antibody, LM609, prior to replating on rTGFBI. Relative adhesion was quantitated by measuring the absorbance at 540 nm and values were normalized to an uncoated well. Results represent 3 independent experiments and error bars represent standard deviation. *p < 0.05, **p < 0.001. (AVI 828 KB)12943_2011_1019_MOESM5_ESM.avi
Additional file 6: Figure S4. Immunofluorescence microscopy of SKOV3 cells stably expressing control or β1 integrin shRNA. Fixed and permeabilized cells were immunostained for paxillin, to highlight focal adhesions, and the actin cytoskeleton was visualized with Alexa Flour 568-phallloidin. Scale bar 40 μm. (AVI )12943_2011_1019_MOESM6_ESM.avi
Authors’ original file for figure 112943_2011_1019_MOESM7_ESM.bmp
Authors’ original file for figure 212943_2011_1019_MOESM8_ESM.bmp
Authors’ original file for figure 312943_2011_1019_MOESM9_ESM.bmp
Authors’ original file for figure 412943_2011_1019_MOESM10_ESM.bmp
Authors’ original file for figure 512943_2011_1019_MOESM11_ESM.bmp
Authors’ original file for figure 612943_2011_1019_MOESM12_ESM.bmp
Authors’ original file for figure 712943_2011_1019_MOESM13_ESM.bmp
Ahmed AA, Mills AD, Ibrahim AE, Temple J, Blenkiron C, Vias M, Massie CE, Iyer NG, McGeoch A, Crawford R: The extracellular matrix protein TGFBI induces microtubule stabilization and sensitizes ovarian cancers to paclitaxel. Cancer Cell. 2007, 12 (6): 514-527. 10.1016/j.ccr.2007.11.014 PubMedCentralCrossRefPubMed
Bull Phelps SL, Carbon J, Miller A, Castro-Rivera E, Arnold S, Brekken RA, Lea JS: Secreted protein acidic and rich in cysteine as a regulator of murine ovarian cancer growth and chemosensitivity. Am J Obstet Gynecol. 2009, 200 (2): e181-187. 180.
Sherman-Baust CA, Weeraratna AT, Rangel LB, Pizer ES, Cho KR, Schwartz DR, Shock T, Morin PJ: Remodeling of the extracellular matrix through overexpression of collagen VI contributes to cisplatin resistance in ovarian cancer cells. Cancer Cell. 2003, 3 (4): 377-386. 10.1016/S1535-6108(03)00058-8 CrossRefPubMed
Calaf GM, Echiburu-Chau C, Zhao YL, Hei TK: BigH3 protein expression as a marker for breast cancer. Int J Mol Med. 2008, 21 (5): 561-568. PubMed
Yamanaka M, Kimura F, Kagata Y, Kondoh N, Asano T, Yamamoto M, Hayakawa M: BIGH3 is overexpressed in clear cell renal cell carcinoma. Oncol Rep. 2008, 19 (4): 865-874. PubMed
Buckhaults P, Rago C, St Croix B, Romans KE, Saha S, Zhang L, Vogelstein B, Kinzler KW: Secreted and cell surface genes expressed in benign and malignant colorectal tumors. Cancer Res. 2001, 61 (19): 6996-7001. PubMed
Hashimoto K, Noshiro M, Ohno S, Kawamoto T, Satakeda H, Akagawa Y, Nakashima K, Okimura A, Ishida H, Okamoto T: Characterization of a cartilage-derived 66-kDa protein (RGD-CAP/beta ig-h3) that binds to collagen. Biochim Biophys Acta. 1997, 1355 (3): 303-314. 10.1016/S0167-4889(96)00147-4 CrossRefPubMed
Kim JE, Park RW, Choi JY, Bae YC, Kim KS, Joo CK, Kim IS: Molecular properties of wild-type and mutant betaIG-H3 proteins. Invest Ophthalmol Vis Sci. 2002, 43 (3): 656-661. PubMed
Kim BY, Olzmann JA, Choi SI, Ahn SY, Kim TI, Cho HS, Suh H, Kim EK: Corneal dystrophy-associated H124H mutation disrupts TGFBIinteraction with periostin and causes mislocalization to the lysosome. J Biol Chem. 2009.
Kawamoto T, Noshiro M, Shen M, Nakamasu K, Hashimoto K, Kawashima-Ohya Y, Gotoh O, Kato Y: Structural and phylogenetic analyses of RGD-CAP/beta ig-h3, a fasciclin-like adhesion protein expressed in chick chondrocytes. Biochim Biophys Acta. 1998, 1395 (3): 288-292. 10.1016/S0167-4781(97)00172-3 CrossRefPubMed
Gillan L, Matei D, Fishman DA, Gerbin CS, Karlan BY, Chang DD: Periostin secreted by epithelial ovarian carcinoma is a ligand for alpha(V)beta(3) and alpha(V)beta(5) integrins and promotes cell motility. Cancer Res. 2002, 62 (18): 5358-5364. PubMed
Horiuchi K, Amizuka N, Takeshita S, Takamatsu H, Katsuura M, Ozawa H, Toyama Y, Bonewald LF, Kudo A: Identification and characterization of a novel protein, periostin, with restricted expression to periosteum and periodontal ligament and increased expression by transforming growth factor beta. J Bone Miner Res. 1999, 14 (7): 1239-1249. 10.1359/jbmr.19126.96.36.1999 CrossRefPubMed
Baril P, Gangeswaran R, Mahon PC, Caulee K, Kocher HM, Harada T, Zhu M, Kalthoff H, Crnogorac-Jurcevic T, Lemoine NR: Periostin promotes invasiveness and resistance of pancreatic cancer cells to hypoxia-induced cell death: role of the beta4 integrin and the PI3k pathway. Oncogene. 2007, 26 (14): 2082-2094. 10.1038/sj.onc.1210009 CrossRefPubMed
Oh JE, Kook JK, Min BM: Beta ig-h3 induces keratinocyte differentiation via modulation of involucrin and transglutaminase expression through the integrin alpha3beta1 and the phosphatidylinositol 3-kinase/Akt signaling pathway. J Biol Chem. 2005, 280 (22): 21629-21637. 10.1074/jbc.M412293200 CrossRefPubMed
Kim JE, Jeong HW, Nam JO, Lee BH, Choi JY, Park RW, Park JY, Kim IS: Identification of motifs in the fasciclin domains of the transforming growth factor-beta-induced matrix protein betaig-h3 that interact with the alphavbeta5 integrin. J Biol Chem. 2002, 277 (48): 46159-46165. 10.1074/jbc.M207055200 CrossRefPubMed
Albini A, Allavena G, Melchiori A, Giancotti F, Richter H, Comoglio PM, Parodi S, Martin GR, Tarone G: Chemotaxis of 3 T3 and SV3T3 cells to fibronectin is mediated through the cell-attachment site in fibronectin and a fibronectin cell surface receptor. J Cell Biol. 1987, 105 (4): 1867-1872. 10.1083/jcb.105.4.1867 CrossRefPubMed
Kalra J, Warburton C, Fang K, Edwards L, Daynard T, Waterhouse D, Dragowska W, Sutherland BW, Dedhar S, Gelmon K, Bally M: QLT0267, a small molecule inhibitor targeting integrin-linked kinase (ILK), and docetaxel can combine to produce synergistic interactions linked to enhanced cytotoxicity, reductions in P-AKT levels, altered F-actin architecture and improved treatment outcomes in an orthotopic breast cancer model. Breast Cancer Res. 2009, 11 (3): R25 10.1186/bcr2252 PubMedCentralCrossRefPubMed
Danen EH, Sonneveld P, Brakebusch C, Fassler R, Sonnenberg A: The fibronectin-binding integrins alpha5beta1 and alphavbeta3 differentially modulate RhoA-GTP loading, organization of cell matrix adhesions, and fibronectin fibrillogenesis. J Cell Biol. 2002, 159 (6): 1071-1086. 10.1083/jcb.200205014 PubMedCentralCrossRefPubMed
Skonier J, Bennett K, Rothwell V, Kosowski S, Plowman G, Wallace P, Edelhoff S, Disteche C, Neubauer M, Marquardt H: beta ig-h3: a transforming growth factor-beta-responsive gene encoding a secreted protein that inhibits cell attachment in vitro and suppresses the growth of CHO cells in nude mice. DNA Cell Biol. 1994, 13 (6): 571-584. 10.1089/dna.1994.13.571 CrossRefPubMed
Andersen RB, Karring H, Moller-Pedersen T, Valnickova Z, Thogersen IB, Hedegaard CJ, Kristensen T, Klintworth GK, Enghild JJ: Purification and structural characterization of transforming growth factor beta induced protein (TGFBIp) from porcine and human corneas. Biochemistry. 2004, 43 (51): 16374-16384. 10.1021/bi048589s CrossRefPubMed
- β3 integrin modulates transforming growth factor beta induced (TGFBI) function and paclitaxel response in ovarian cancer cells
David A Tumbarello
James D Brenton
- BioMed Central
Neu im Fachgebiet Onkologie
Mail Icon II