PMID- 19405959 OWN - NLM STAT- MEDLINE DCOM- 20090811 LR - 20231213 IS - 1465-542X (Electronic) IS - 1465-5411 (Print) IS - 1465-5411 (Linking) VI - 11 IP - 2 DP - 2009 TI - Tumour-associated tenascin-C isoforms promote breast cancer cell invasion and growth by matrix metalloproteinase-dependent and independent mechanisms. PG - R24 LID - 10.1186/bcr2251 [doi] AB - INTRODUCTION: The stromal microenvironment has a profound influence on tumour cell behaviour. In tumours, the extracellular matrix (ECM) composition differs from normal tissue and allows novel interactions to influence tumour cell function. The ECM protein tenascin-C (TNC) is frequently up-regulated in breast cancer and we have previously identified two novel isoforms - one containing exon 16 (TNC-16) and one containing exons 14 plus 16 (TNC-14/16). METHODS: The present study has analysed the functional significance of this altered TNC isoform profile in breast cancer. TNC-16 and TNC-14/16 splice variants were generated using PCR-ligation and over-expressed in breast cancer cells (MCF-7, T47D, MDA-MD-231, MDA-MB-468, GI101) and human fibroblasts. The effects of these variants on tumour cell invasion and proliferation were measured and compared with the effects of the large (TNC-L) and fully spliced small (TNC-S) isoforms. RESULTS: TNC-16 and TNC-14/16 significantly enhanced tumour cell proliferation (P < 0.05) and invasion, both directly (P < 0.01) and as a response to transfected fibroblast expression (P < 0.05) with this effect being dependent on tumour cell interaction with TNC, because TNC-blocking antibodies abrogated these responses. An analysis of 19 matrix metalloproteinases (MMPs) and tissue inhibitor of matrix metalloproteinases 1 to 4 (TIMP 1 to 4) revealed that TNC up-regulated expression of MMP-13 and TIMP-3 two to four fold relative to vector, and invasion was reduced in the presence of MMP inhibitor GM6001. However, this effect was not isoform-specific but was elicited equally by all TNC isoforms. CONCLUSIONS: These results demonstrate a dual requirement for TNC and MMP in enhancing breast cancer cell invasion, and identify a significant role for the tumour-associated TNC-16 and TNC-14/16 in promoting tumour invasion, although these isoform-specific effects appear to be mediated through MMP-independent mechanisms. FAU - Hancox, Rachael A AU - Hancox RA AD - Department of Cancer Studies and Molecular Medicine, Infirmary Close, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester, UK. raa9@le.ac.uk FAU - Allen, Michael D AU - Allen MD FAU - Holliday, Deborah L AU - Holliday DL FAU - Edwards, Dylan R AU - Edwards DR FAU - Pennington, Caroline J AU - Pennington CJ FAU - Guttery, David S AU - Guttery DS FAU - Shaw, Jacqueline A AU - Shaw JA FAU - Walker, Rosemary A AU - Walker RA FAU - Pringle, J Howard AU - Pringle JH FAU - Jones, J Louise AU - Jones JL LA - eng GR - 2001:232/BCN_/Breast Cancer Now/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20090430 PL - England TA - Breast Cancer Res JT - Breast cancer research : BCR JID - 100927353 RN - 0 (Protein Isoforms) RN - 0 (RNA, Messenger) RN - 0 (TIMP3 protein, human) RN - 0 (Tenascin) RN - 0 (Tissue Inhibitor of Metalloproteinase-1) RN - 0 (Tissue Inhibitor of Metalloproteinase-3) RN - 0 (Tissue Inhibitor of Metalloproteinases) RN - 127497-59-0 (Tissue Inhibitor of Metalloproteinase-2) RN - EC 3.4.24.- (Matrix Metalloproteinases) SB - IM MH - Alternative Splicing MH - Blotting, Western MH - Breast Neoplasms/*enzymology/genetics/*pathology MH - Cell Adhesion MH - Cell Movement MH - Cell Proliferation MH - Female MH - Humans MH - Matrix Metalloproteinases/genetics/*metabolism MH - Neoplasm Invasiveness MH - Protein Isoforms MH - RNA, Messenger/genetics/metabolism MH - Reverse Transcriptase Polymerase Chain Reaction MH - Tenascin/*physiology MH - Tissue Inhibitor of Metalloproteinase-1/genetics/metabolism MH - Tissue Inhibitor of Metalloproteinase-2/genetics/metabolism MH - Tissue Inhibitor of Metalloproteinase-3/genetics/metabolism MH - Tissue Inhibitor of Metalloproteinases/genetics/metabolism MH - Up-Regulation MH - Tissue Inhibitor of Metalloproteinase-4 PMC - PMC2688953 EDAT- 2009/05/02 09:00 MHDA- 2009/08/12 09:00 PMCR- 2009/04/30 CRDT- 2009/05/02 09:00 PHST- 2008/08/04 00:00 [received] PHST- 2009/01/15 00:00 [revised] PHST- 2009/04/30 00:00 [accepted] PHST- 2009/05/02 09:00 [entrez] PHST- 2009/05/02 09:00 [pubmed] PHST- 2009/08/12 09:00 [medline] PHST- 2009/04/30 00:00 [pmc-release] AID - bcr2251 [pii] AID - 10.1186/bcr2251 [doi] PST - ppublish SO - Breast Cancer Res. 2009;11(2):R24. doi: 10.1186/bcr2251. Epub 2009 Apr 30.