Abstract
BackgroundZedoary (Curcumae Rhizoma, Ezhu), a Chinese medicinal herb, has been reported to show anticancer activity. This study aims to investigate the effect of zedoary oil (Ezhu You) on the proliferation of AGS cells which is one gastric cancer cell line.MethodsThe main ingredients of the herb were detected by GC-MS for herbal quality control. Cell viability was measured by MTT assay and cell proliferation was investigated by immunocytochemical staining for proliferating cell nuclear antigen (PCNA) protein. In addition, the cell cycle distributions were detected by flow cytometry with propidium iodine (PI) staining and the apoptosis rates were evaluated by flow cytometry with annexin V/PI double-staining. The morphological changes associated with apoptosis were observed by Hoechst 33342/PI double-staining. Protein expression was determined by western blot analysis.ResultsThe main ingredients of the herb, including curzerene (26.45%), eucalyptol (12.04%), curcumol (9.04%), pyridine (7.97%), germacrone (7.89%), β-elemene (7.36%), τ-elemene (4.11%) and 28 other ingredients, including curdione, were consistent with the chemical profiles of zedoary. Zedoary oil significantly decreased the cell viability of AGS cells (P < 0.01) and MGC 803 cells (P < 0.01), and the inhibitory effects were attenuated by elevated concentrations of FBS. At high concentrations (≥90 μg/mL), zedoary oil killed GES-1 cells. At low concentrations (≤60 μg/mL), zedoary oil was less inhibitory toward normal gastric epithelial cells than gastric cancer cell lines. In AGS cells, zedoary oil inhibited cell proliferation in a dose- and time-dependent manner, with decreased PCNA protein expression in the zedoary oil-treated cells, and arrested the cell cycle at S, G2/M and G0/G1 stages after treatment for 6–48 h. At concentrations of 30, 60 and 90 μg/mL, which resulted in significant inhibition of proliferation and cell cycle arrest, zedoary oil induced cell apoptosis. In addition, Hoechst 33342/PI double-staining confirmed the morphological characteristics of cell apoptosis at 24 h. Zedoary oil upregulated the ratio of Bax/Bcl-2 protein expression (P < 0.01).ConclusionsZedoary oil inhibited AGS cell proliferation through cell cycle arrest and cell apoptosis promotion, which were related to Bax/Bcl-2 protein expression.
Highlights
Zedoary (Curcumae Rhizoma, Ezhu), a Chinese medicinal herb, has been reported to show anticancer activity
Many other compounds were detected by the gas chromatography–mass spectrometry (GC-MS), and several chemical compounds could not be determined from the National Institute of Standards and Technology (NIST) library
Inhibitory effects of zedoary oil on cell viability Zedoary oil inhibited the proliferation of AGS cells in a dose- and time-dependent manner after treatment for 24, 48 and 72 h (P < 0.01 vs. control cells) (Table 2)
Summary
Zedoary (Curcumae Rhizoma, Ezhu), a Chinese medicinal herb, has been reported to show anticancer activity. This study aims to investigate the effect of zedoary oil (Ezhu You) on the proliferation of AGS cells which is one gastric cancer cell line. Curcuma kwangsiensis and Curcuma wenyujin are named zedoary in the Chinese Pharmacopoeia and used as antiviral and antimicrobial medicines [1,2,3]. Zedoary oil is a Chinese medicine that is used for treatment of gynecologic inflammation [4], pneumonia [5], pediatric diseases [6], viral myocarditis [7] and malignant tumors, such as oophoroma, hepatocellular carcinoma and lung. Gastric glandular cells are replaced by intestinal-type epithelial cells with high cell proliferation rates and fibrosis in severe inflammation of the gastric membrane, resulting in chronic atrophic gastritis with intestinal metaplasia and dysplasia, imbalance between cell proliferation and apoptosis in the normal gastric mucosa and increasing incidence of gastric cancer [16,17,18,19].
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