Antcin-H Isolated from Antrodia cinnamomea Inhibits Renal Cancer Cell Invasion Partly through Inactivation of FAK-ERK-C/EBP-β/c-Fos-MMP-7 Pathways

Figure 6

Inhibition of c-Fos and C/EBP- activities involved in antcin-H-mediated MMP-7 downregulation. (a) Reporter luciferase assay. 786-0 cells were transiently transfected with reporter vector containing MMP-7 promoter +1−1500 region or control vector for 24 h, and then the cells were treated without or with 20, 50, and 100 M antcin-H for another 24 h. After incubation, the luciferase activity was measured and the relative luciferase activity was presented as means ± SD. Statistically significant, , , and . (b) Putative binding sites of c-Fos and C/EBP- located at upstream of MMP-7 promoter. (c) Antcin-H prevents c-Fos and C/EBP- binding to the MMP-7 upstream promoter/response element region. The 786-0 cells were treated without or with 100 μM antcin-H for 24 h, the cells were collected, and then ChIP assay was carried out. (d) Antcin-H dose-dependently inhibits c-Fos and C/EBP- binding to the MMP-7 upstream promoter/response element region. The 786-0 cells were treated without or with 20, 50, and 100 μM antcin-H for 24 h, the cells were collected, and then the activity of c-Fos and C/EBP- binding to each response site located at MMP-7 promoter upstream was determined by ChIP assay.

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