Potential Therapeutic Role of Z-Isochaihulactone in Lung Cancer through Induction of Apoptosis via Notch Signaling
Figure 2
Z-isochaihulactone caused growth inhibition by inducing G2/M phase arrest of human lung adenocarcinoma cells in vitro and induces apoptosis. (a) Growth inhibition effect was assessed in human lung cancer cell lines A549. The cells were treated with various concentrations ranging from 1.25 to 20 μM Z-isochaihulactone for various times (24, 48, and 72 h) as indicated. Growth inhibition effect (IC50) was determined by MTT assay. (b) Cells were treated with 10 μM Z-isochaihulactone for 48 h then stained with Hoechst 33342 for 30 min. The chromatin morphology of A549 cells was measured by fluorescence microscopy (magnification 400x). (c) The Western blot analysis was done and as described in Section 2. Cells were treated with Z-isochaihulactone with various concentrations ranging from 1.25–20 μM for 24 h. Cells were probed with Cleaved caspase-9, caspase-9, Cleaved caspase-3, caspase-3, Cleaved PARP, and PARP antibodies. Expression of β-actin was used as an internal control. (d) Cells were treated with 5 μM Z-isochaihulactone for the indicated time (6–48 h) and analyzed by Western blot analysis. Cells were probed with Cleaved caspase-9, caspase-9, Cleaved caspase-3, caspase-3, Cleaved PARP, and PARP antibodies. Expression of β-actin was used as an internal control. (e) The cell cycle analysis was done as described in Section 2. The cell cycle was assessed in A549 cells. Cells were treated with various concentrations from 2.5 to 10 μM Z-isochaihulactone and VPA 10 mM for 48 h. Cells were collected, fixed in alcohol, then stained with propidium iodide (PI), and analyzed by flow cytometry. (f) Cells were treated with various concentrations of Z-isochaihulactone from 1.25 to 20 μM and VPA 10 mM for 48 h. The cell cycle association proteins were determined by Western blot analysis. The Western blot analysis was done and as described in Section 2. Cells were probed with cyclin B1 and phosphor-cdc2 (Thr161) antibodies. Expression of β-actin was used as an internal control.
