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. 2011 Jan 13;6(1):e15313. doi: 10.1371/journal.pone.0015313

Figure 2. Breast cancer-associated fibroblasts expressed higher HGF levels than normal tissue-associated fibroblasts.

Figure 2

(A) Regression of the cytokine/growth factor signal intensities in the conditional media from CAF #199C and NAF #200N was performed to evaluate the difference of the cytokines and growth factors secreted by CAFs and NAFs. The results revealed that HGF (4.57 fold) and TIMP-1 (1.41 fold) levels were significantly higher in the conditional medium from CAFs than those in the conditional medium from NAFs (array B). In contrast, lower levels of GCP-2 (0.17 fold) (array A), IGFBP-3 (0.48 fold), GRO family (0.44 fold), GRO-α (0.6 fold), ENA-78 (0.63 fold), GCSF (0.54 fold) (array B) and LAP (0.59 fold) (array C) were detected in the conditional media from CAFs compared to those in the conditional medium from NAFs. (B) Western blotting analysis revealed that the HGF protein levels in CAFs were higher than those in NAFs in all fibroblast pairs (upper panel). Whereas, there was no correlation observed between CAFs and IGFBP-3 protein expression (lower panel). (C) Enzyme-linked immunosorbent assay showed that the HGF amounts in the cultured media of CAFs were higher than those of NAFs in all fibroblast pairs. (D) Quantitative real-time RT-PCR analysis showed that the HGF mRNA levels in CAFs were higher than those in NAFs in all fibroblast pairs. (E) Real-time RT-PCR analysis revealed that the mRNA levels of SDF-1 α, SDF-1 β and SDF-1 γ varied in different samples. There was no association between the SDF-1 mRNA levels and CAFs. Data are mean ± SD of triplicate samples. * P<0.05. NS, no significant difference.