羧基-D-氨基葡萄糖对人食管癌Eca-109细胞caspase-3活化及bcl-2蛋白表达的影响

Effect of 2-((3-carboxy-1-oxopropyl)amino)-2-deoxy-D-glucose on Caspase-3 activation and Bcl-2 expression in human esophageal cancer cell line Eca-109

目的探讨D-氨基葡萄糖衍生物2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖(COPADG)对人食管癌Eca-109细胞凋亡的影响及其作用机制。方法体外培养Eca-109细胞;倒置显微镜下观察细胞形态学变化;流式细胞术检测COPADG作用后Eca-109细胞内半胱氨酸酶3(caspase-3)活化程度及bcl-2蛋白表达的变化;Annexin V/PI染色结合流式细胞术检测COPADG作用后Eca-109细胞凋亡率。结果COPADG作用显著增加Eca-109细胞凋亡率,且Eca-109细胞内caspase-3活性和bcl-2表达平均荧光强度明显增高。结论COPADG能显著诱导Eca-109细胞凋亡发生,并使Eca-109细胞内caspase-3显著活化以及bcl-2蛋白表达下调,并可能通过该机制诱导细胞凋亡的发生。

Objective To observe the effect of D-glucosamine 2-（（3-carboxy-1-oxopropyl） amino）-2-deoxy-D-glucose （COPADG） on apoptosis and caspase-3 activation and bcl-2 protein expression in human esophageal cancer Eca-109 cells and discuss the possible mechanisms. Methods Eca-109 cells were cultured in vitro, then exposured to 0.03 mmol/L COPADG for different time periods. The cell morphological changes were observed by inverted phase contrast microscopy. Caspase-3 activity and bcl-2 expression were detected by flow eytometry, and apoptosis rate was analyzed by Annexin V/PI fluorescence staining together with flow cytometry. Results COPADG could significantly induce Eca-109 cells to apoptosis, and increase mean fluorescence intensity of caspase-3 and bcl-2 protein expressions in the cells. Conclusion COPADG induces the apoptosis of human esophageal cancer cell line Eca-109, as well as the activation of caspase-3 and the downregulated expression of bcl-2 protein. This may not an unique mechanism in COPADG-induced apoptosis.