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Effect of Water Extracts from Dendrobium huoshanense on Proliferation and Apoptosis of Human Laryngeal Carcinoma Hep-2 Cells and Relevant Mechanism

ZHANG Jing△,LIAN Chao-qun,WU Shou-wei,HU Ming-jie,CHEN Chuan-hao(△Bioscience Department,Bengbu Medical College,Bengbu 233030,Anhui Province,China)  
Objective To investigate the effect of water extracts from Dendrobium huoshanense(WEDH) on proliferation and apoptosis of human laryngeal carcinoma Hep-2 cells and the relevant mechanism.Methods Hep-2 cells were treated with 3.75,7.50,15.00,30.00 and 60.00 mg / ml WEDH respectively and observed for proliferation level by MTT method,for morphology by invert microscopy and for apoptosis by laser scanning confocal microscopy after stained with Annexin V-FITC / PI,then determined for free calcium ion(i) concentration by laser scanning confocal microscopy after stained with Fluo-3 AM and for caspase-3 activity by spectrophotometry.Results WEDH showed significantly time-and dose-dependent inhibitory effect on proliferation of Hep-2 cells.The IC50 of WEDH to Hep-2 cells 24,48 and 72 h after treatment were 48.54,26.67 and 14.93 mg / ml respectively.The morphology of Hep-2 cells 48 h after treatment with WEDH changed significantly,in a dose-dependent mode,and showed typical characteristics of apoptosis.Bothi concentration and caspase-3 activity of Hep-2 cells were significantly higher than those in negative control group(P 0.05).Conclusion WEDH inhibited the proliferation and induced the apoptosis of Hep-2 cells significantly by a potential mechanism which might be associated with increasing the i concentration and activating caspase-3.
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