A New Herbal Formula, KSG-002, Suppresses Breast Cancer Growth and Metastasis by Targeting NF-κB-Dependent TNFα Production in Macrophages
Figure 4
KSG-002 inhibits TNFα production. (a) TNFα levels were analyzed with ELISA assays. Experiments were performed in triplicate. Bars indicate means ± standard deviations. . (b) KSG-002 inhibits NF-κB signaling pathway. Western blots were performed with the appropriate antibodies. (c) KSG-002 inhibits expressions of NF-κB-dependent genes. Western blots were done using antibodies for proteins indicated. Tubulin was used for the loading control. (d) The luciferase activities. Raw264.7 cells were transfected with p65 plasmid and NF-κB-luc (top) or TNFα-luc (bottom) plasmid for 24 hours and then treated with KSGs (200 μg/mL) for another 6 hours. The experiments were performed in triplicate. . (e) MDA-MB-231 cells were scratched and treated with 100 ng/mL of TNFα alone or cotreated with TNFα (102 ng/mL, colored in gray) and KSGs for 24 hours. Right triangles indicate different concentrations of KSGs (50, 100, 200, and 500 μg/mL). Migrating cell numbers were counted, and data were presented as means ± standard deviations. Experiments were performed three times. , TNF alone versus TNFα plus KSG-002. (f and g) Cells indicated were treated with TNFα alone (f) or together with KSGs (g) for 48 hours and subjected to MTT assays. Right triangles indicate different concentrations of TNFα (10−3, 10−2, 10−1, 1, 10, and 102 ng/mL). For the combination, 102 ng/mL of TNFα was treated with different concentrations of KSGs as indicated. Experiments were performed three times in triplicate. Representative data show mean ± standard deviation. .
