TLR4 Ligand/H2O2 Enhances TGF-β1 Signaling to Induce Metastatic Potential of Non-Invasive Breast Cancer Cells by Activating Non-Smad Pathways
Figure 1
TGF-β1/H2O2/LPS facilitates invasive capability of non-invasive breast cancer cells.
(A) MCF-7 and T-47D cells were cultured in absence or presence of TGF-β1 (5 ng/ml)/H2O2 (50 µM)/LPS (100 ng/ml) for the indicated time, and then used for Matrigel invasion assay. (B–D) Tumor cells were cultured in absence or presence of TGF-β1, H2O2 and LPS for 8 days. The cells were then used for following experiments. (B) The cells were used for Matrigel invasion assay. (C) The expression of αvβ3 was analyzed by real-time RT-PCR and flow cytometry. αvβ3 expression index was calculated as described in Materials and Methods. (D) The cells were then cultured in presence of matrigel for 48 h. The mRNA level of MMP9 was detected by real-time RT-PCR. The MMP-9 in supernatants was detected by zymography assay, and the fold difference of active MMP-9 was calculated after densitometric analysis of the gel. P values, *P<0.05, **P<0.01.