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
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- 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
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