Potential Therapeutic Role of Z-Isochaihulactone in Lung Cancer through Induction of Apoptosis via Notch Signaling
Figure 3
Effect of Z-isochaihulactone on protein expression of notch signaling association gene and induction of apoptosis in A549 cells. (a) Cells were treated with various concentrations of Z-isochaihulactone from 1.25 to 202 μM and VPA 10 mM for 48 h (upper panel) or treated with 5 μM Z-isochaihulactone for various times (6–48 h). (b) The activated Notch1 (Notch1 intracellular domain, NICD) protein expression was measured by Western blot. (c) Cells were treated with various concentrations of Z-isochaihulactone from 1.25 to 20 μM and VPA 10 mM for 48 h or treated with 5 μM Z-isochaihulactone for various times (6–48 h). The notch signaling pathway and apoptosis association protein expressions (p21, phosphor-c-Myc, NF-κB, and Hes1) were measured. The Western blot analysis was done and as described in Section 2. Expression of β-actin was used as an internal control. (d) A549 cells were transfected with pEF.hICN1.CMV.GFP (human intracellular Notch1 gene) using GeneJammer transfection reagent. The gene expression was measured by fluorescence microscopy. (e) Inhibition of Notch1 expression and growth inhibition by γ-secretase inhibitor L-685. The Notch1 expression of A549 cells with culture or serum-containing medium was pretreated with the γ-secretase inhibitor L-685 for 1 hr and then treated with 5 μM Z-isochaihulactone for various times (6–48 h), and cell apoptosis percentage was measured using flow cytometry. Each column represents the mean ± SD (; ).
