Figure 5

Effect of fibroblast-associated tenascin-C expression on MCF-7 tumour cell invasion. (a) Mean invasion index (MII) for MCF-7 tumour cell co-cultured with primary breast fibroblasts (n = 5) transfected with tenascin-C (TNC) isoforms constructs or vector alone. Fibroblasts expressing TNC-16 or TNC-14/16 significantly increased MII compared with TNC-L (P = 0.05 and P = 0.001, respectively), TNC-S and vector alone (***P < 0.001). Higher MII was also seen with fibroblast expressing TNC-L (P < 0.001) compared with TNC-S and vector alone. Bars indicate the mean of five donors, each measured in triplicate. (b) MCF-7 cell MII using conditioned media from primary breast fibroblasts transfected with TNC isoforms constructs or the vector alone in the presence of blocking TNC mouse monoclonal antibody BC-24 or equivalent IgG control. The blocking antibody significantly reduced MII (*P < 0.05) independent of the TNC isoform. In all cases, the bars indicate the mean of three experiments with standard errors shown. (c) GI101 cell MII using conditioned media from fibroblasts transfected with TNC isoforms constructs or the vector alone in the presence of blocking TNC mouse monoclonal antibody BC-24 or equivalent IgG control. The blocking antibody significantly reduced MII (*P < 0.05, **P < 0.01) in all the cases except for TNC-16. In all cases, the bars indicate the mean of three experiments with standard errors shown. (d) MCF-7 cells transfected with different TNC isoforms or vector only all exhibited significant reduction in MII in the presence of the matrix metalloproteinase inhibitor GM6001 (***P < 0.001). Bars indicate mean of three experiments.