Evidence-Based Complementary and Alternative Medicine / 2013 / Article / Fig 5

Research Article

Honokiol Eliminates Human Oral Cancer Stem-Like Cells Accompanied with Suppression of Wnt/β-Catenin Signaling and Apoptosis Induction

Figure 5

Honokiol downregulated the Wnt/β-catenin signaling and its downstream in SAS SP cells. (a) Honokiol decreased the protein levels of β-catenin and TCF-4 as well as their downstream c-Myc and Cyclin D1, accompanied with the increase of GSK-3α/β in SP cells. The expression of stem cell marker CD44 was also decreased. Cells were treated with honokiol for 48 h and the cell lysates were analyzed with Western blot using β-actin as loading control. (b) TOPFLASH TCF reporter assay in honokiol-treated SP cells. Cells were transfected with pTOPFLASH luciferase reporter constructs as described in Section 2 and treated with honokiol for 48 h, lysed, and analyzed using the Dual Luciferase Assay System kit. Results are expressed in relative luciferase units (RLU), normalized to transfection efficiency as described in Section 2. Each bar represents the mean of three independent transfections ±S.E. (c) Honokiol downregulated the EMT markers Slug and Snail proteins as well as Cyclin D1 while suppressed the β-catenin in SP cells. Cells were treated with honokiol for 48 h and then analyzed by Western blot as described in (a).

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