Figure 2

Induction of apoptosis in NSCLC (H1299 and A549) cell lines, normal bronchial epithelial (Beas2b) cells and human fibroblasts (Wi38). (A) Apoptosis was evaluated as described in Materials and Methods with Annexin V staining in H1299, A549, Beas2B and Wi38 cells after treatment with (60 μg/mL for 72 h) and without (CTR-) Urtica dioica extract. Representative dot plot diagrams of flow cytometric analysis of H1299 and Wi38 cell apoptosis are shown. Dot plot diagrams show the different stages of apoptosis. % indicated in the UL (Upper Left) quadrant represent cells positive for Annexin V and negative for 7AAD, considered as early apoptotic cells; % in UR (Upper Right) quadrant indicate cells positive for both Annexin V and 7AAD, showing the late apoptotic or necrotic cells population; % in LL (Lower Left) quadrant are negative for both markers and represent viable cells. Histogram of data expressed as percentage of both early and late apoptotic cells. Bars represent mean values obtained from three separate experiments. P values < 0.05 were considered as statistically significant (**) at Student-t test. (B) The Western Blot analyses, which were carried out using antibodies against PARP, (89)-cleaved-PARP fragment, pro-caspase3, pro-caspase 8 and cleaved BID, were performed on protein lysates from cell after the indicated treatment with (+) and without (−) Urtica dioica extract.