Plumbagin from Plumbago Zeylanica L Induces Apoptosis in Human Non-small Cell Lung Cancer Cell Lines through NF-κB Inactivation

Tong-Peng Xu, Hua Shen, Ling-Xiang Liu, Yong-Qian Shu
2013 Asian Pacific Journal of Cancer Prevention  
Objective: To detect effects of plumbagin on proliferation and apoptosis in non-small cell lung cancer cell lines, and investigate the underlying mechanisms. Materials and Methods: Human non-small cell lung cancer cell lines A549, H292 and H460 were treated with various concentrations of plumbagin. Cell proliferation rates was determined using both cell counting kit-8 (CCK-8) and clonogenic assays. Apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry and TUNEL
more » ... . The levels of reactive oxygen species (ROS) were detected by flow cytometry. Activity of NF-kB was examined by electrophoretic mobility shift assay (EMSA) and luciferase reporter assay. Western blotting was used to assess the expression of both NF-kB regulated apoptotic-related gene and activation of p65 and IkBk. Results: Plumbagin dose-dependently inhibited proliferation of the lung cancer cells. The IC50 values of plumbagin in A549, H292, and H460 cells were 10.3 μmol/L, 7.3 μmol/L, and 6.1 μmol/L for 12 hours, respectively. The compound concentration-dependently induced apoptosis of the three cell lines. Treatment with plumbagin increased the intracellular level of ROS, and inhibited the activation of NK-kB. In addition to inhibition of NF-kB/p65 nuclear translocation, the compound also suppressed the degradation of IkBk. ROS scavenger NAC highly reversed the effect of plumbagin on apoptosis and inactivation of NK-kB in H460 cell line. Treatment with plumbagin also increased the activity of caspase-9 and caspase-3, downregulated the expression of Bcl-2, upregulated the expression of Bax, Bak, and CytC. Conclusions: Plumbagin inhibits cell growth and induces apoptosis in human lung cancer cells through an NF-kB-regulated mitochondrial-mediated pathway, involving activation of ROS.
doi:10.7314/apjcp.2013.14.4.2325 fatcat:jaqsllkqi5bjzktd5nzdtybfcu