Synergistic apoptotic effect of the combination of diosgenin and TRAIL on non-small-cell lung cancer cell line A549 evaluated with the Chou-Talalay method.
- Author:
Yan HE
1
;
Ji-Shuang WANG
;
Peng ZHANG
;
Wen-Jing ZHANG
;
Qi-Lai HUANG
;
Zi-Chun HUA
Author Information
1. Faculty of Traditional Chinese Medicine and Macau Institute for Applied Research in Medicine and Health, the State Key Laboratory for Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau 999078, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
drug effects;
Apoptosis Regulatory Proteins;
metabolism;
Carcinoma, Non-Small-Cell Lung;
metabolism;
pathology;
Cell Line, Tumor;
Cell Proliferation;
drug effects;
Diosgenin;
administration & dosage;
pharmacology;
Dose-Response Relationship, Drug;
Drug Synergism;
HEK293 Cells;
Humans;
Lung Neoplasms;
metabolism;
pathology;
Mitogen-Activated Protein Kinase Kinases;
metabolism;
Signal Transduction;
TNF-Related Apoptosis-Inducing Ligand;
administration & dosage;
pharmacology
- From:
Acta Pharmaceutica Sinica
2013;48(1):45-51
- CountryChina
- Language:Chinese
-
Abstract:
This study is to investigate the apoptotic induction effect of the combination of diosgenin and TNF-related apoptosis-inducing ligand (TRAIL) on non-small-cell lung cancer cell line A549 by using the Chou-Talalay method, and observe the mechanism of the combination. The apoptotic effect of diosgenin or TRAIL alone and their combination on A549 and normal cell line 293T proliferation was measured by MTT assay. Chou-Talalay method was used to evaluate the combination effect. Apoptosis was examined by Hoechst 33342 staining and flow cytometry assay. Western blotting detects the expression of apoptosis-associated proteins. Diosgenin or TRAIL alone can inhibit proliferation ofA549 in a concentration-dependent manner. According to the Chou-Talalay method, when f(a) = 0.1, CI > 1, when f(a) > 0.1, CI < 1. Combined with TRAIL, the IC50 of diosgenin decreases from 21.864 to 14.810 micromol x L(-1) (P < 0.05) on A549 cells. But for 293T cells, IC50 of diosgenin does not change significantly. As with Hoechst 33342 staining and flow cytometry assay, the apoptosis ratios also increased in the combination group. At protein expression level, combination-treated group displays increased Caspase-8, Caspase-9, Bid, Caspase-3 activation and PARP cleavage, significantly decreased Bcl-2 and increased Bax expression, and MAPK pathways were activated. The combination of diosgenin and TRAIL has synergistic effect on A549 cells.
