Journal: Pharmaceutics

Article Title: Licochalcone A Suppresses Renal Cancer Cell Proliferation and Metastasis by Engagement of Sp1-Mediated LC3 Expression

doi: 10.3390/pharmaceutics15020684

Figure Lengend Snippet: LicA inhibits metastasis via the FAK/Src signaling pathway of RCC cells. Different LicA concentrations (0, 6.25, 12.5, and 12.5 µM) were applied to ACHN cells, and the cells were then left to react for 24 h. ( A ) Western blotting was utilized to examine the expression of p-FAK, t-FAK, p-Src, and t-Src in cell lysates. β-actin served as a internal control. The ACHN cells were treated with 12.5 µM LicA in the presence or absence of ( B ) a FAK inhibitor (PF) or ( C ) an Src inhibitor (PP2) for 24 h. The cell lysates were examined using Western blotting to measure the protein levels of p-FAK, t-FAK, and LC3B. ( D , E ) Next, migration and Matrigel-invasion assays were used to assess migration and invasion. Data represent the mean ± SD of at least three separate experiments. ** p < 0.01, compared to un-treated control (0 µM); # p < 0.05, compared with LicA-treated alone.

Article Snippet: Anti-phosphoryl-FAK (p-FAK), total-FAK (t-FAK), phosphoryl-Src (p-Src) and total-Src (t-Src) antibodies were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Western Blot, Expressing, Migration

Journal: Pharmaceutics

Article Title: Licochalcone A Suppresses Renal Cancer Cell Proliferation and Metastasis by Engagement of Sp1-Mediated LC3 Expression

doi: 10.3390/pharmaceutics15020684

Figure Lengend Snippet: LicA inhibits RCC tumorigenesis and metastasis via the downregulation of the phosphoryl-Src/-FAK pathway. It does so by decreasing the Sp1 transcription activity, resulting in suppressed LC3B expression.

Article Snippet: Anti-phosphoryl-FAK (p-FAK), total-FAK (t-FAK), phosphoryl-Src (p-Src) and total-Src (t-Src) antibodies were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Activity Assay, Expressing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: 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

doi: 10.1155/2017/5052870

Figure Lengend Snippet: Suppression of Src, FAK, and ERK1/2 signaling pathways by antcin-H. (a) Time course-dependent experiment. 786-0 cells were treated without or with 100 μ M antcin-H for 4, 8, and 24 h. (b) Dose-dependent experiment. Cells were treated without or with 20, 50, and 100 μ M antcin-H for 24 h. After incubation, total protein lysates were isolated; the Western blotting analysis was performed to examine the levels of phosphorylated-Src, phosphorylated-FAK, phosphorylated-ERK1/2, phosphorylated-C/EBP- β , and c-Fos. β -Actin was used as an internal loading control.

Article Snippet: Immunoblotting was performed using primary antibodies against Src, phosphorylated-Src (p-Src), paxillin, and phosphorylated-paxillin (p-PXN) (Cell Signaling Technology, Beverly, MA) and ERK1/2, phosphorylated-ERK1/2 (p-ERK1/2), c-Fos, C/EBP- β , phosphorylated-C/EBP- β (p-C/EBP- β ), FAK, and phosphorylated-FAK (p-FAK) (Santa Cruz, CA, USA).

Techniques: Incubation, Isolation, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: 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

doi: 10.1155/2017/5052870

Figure Lengend Snippet: Schematic model of the proposed signaling pathways involved in suppressing cell migration and invasion by antcin-H in human RCC 786-0 cells. Antcin-H inhibits Src, FAK, and ERK1/2 phosphorylated activation, in turn decreasing paxillin, c-Fos, and C/EBP- β activities, reducing the binding of c-Fos and phosphorylated-C/EBP- β to AP-1 and C/EBP- β response elements, thereby decreasing MMPs gene expression, especially MMP-7. Downregulation of MMP-7 and upregulation of TIMP-3 and TIMP-4 gene expression block the degradation of the extracellular matrix proteins and impair the cell invasion. Besides, reducing paxillin phosphorylation and vimentin expression prevents 786-0 cell motility.

Article Snippet: Immunoblotting was performed using primary antibodies against Src, phosphorylated-Src (p-Src), paxillin, and phosphorylated-paxillin (p-PXN) (Cell Signaling Technology, Beverly, MA) and ERK1/2, phosphorylated-ERK1/2 (p-ERK1/2), c-Fos, C/EBP- β , phosphorylated-C/EBP- β (p-C/EBP- β ), FAK, and phosphorylated-FAK (p-FAK) (Santa Cruz, CA, USA).

Techniques: Migration, Activation Assay, Binding Assay, Expressing, Blocking Assay