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Figure 6 | BMC Cancer

Figure 6

From: TGF-β1 modulates the homeostasis between MMPs and MMP inhibitors through p38 MAPK and ERK1/2 in highly invasive breast cancer cells

Figure 6

Relative mRNA and protein expression of MMPs and MMPs inhibitors in MDA-MB-231 cells treated with TGF-β1 and an ERK1/2 inhibitor. MDA-MB-231 cells were pre-treated for 1 h with different concentrations (0, 5, 10 or 20 μM) of PD98059 (ERK1/2 pharmacological inhibitor) and then stimulated with 10 ng/mL TGF-β1 by 20 h. Total RNA from these samples was used to analyze the mRNA expression levels of MMPs (MMP-2 and MMP-9) and MMPs inhibitors (TIMP-2 and RECK) by qRT-PCR. The levels of pro-enzyme and active MMP-2 and MMP-9 proteins were evaluated by zymography. Total protein lysates were used to measure the protein expression levels of RECK by Western blotting. The GAPDH protein expression was used as the loading control in Western blotting assays. Conditioned medium from these cultures were also utilized to analyze the TIMP-2 protein levels by Western blotting. Results are presented in graphics as means ± standard errors from three independent experiments. The Western blotting figures are representative of one experiment. For mRNA expression levels fold change: MMP-2 (a versus c: p < 0.05, a versus d: p < 0.01, a versus e, f: p < 0.001), MMP-9 (a versus c, d, f: p < 0.01), TIMP-2 (a versus b: p < 0.01, a versus c: p < 0.05, a versus d, e, f: p < 0.001) and RECK (a versus b, c: p < 0.05, a versus d, e, f: p < 0.001). For protein expression levels fold change: MMP-2 (a versus c: p < 0.01, a versus d, e: p < 0.05), MMP-9 (a versus c: p < 0.001, c versus f: p < 0.001), TIMP-2 (a versus c: p < 0.01, c versus f: p < 0.001) and RECK (a versus c: p < 0.01, a versus f: p < 0.05, c versus e, f:: p < 0.001).

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