二氢杨梅素诱导人肺腺癌细胞系AGZY-83-a凋亡的实验研究

Dihydromyricetin induce apoptosis in AGZY-83-a tumor cells

目的研究二氢杨梅素(dihydromyricetin)对人肺腺癌细胞系AGZY-83-a凋亡的影响及其相关机制。方法采用MTT法检测二氢杨梅素对人肺腺癌细胞AGZY-83-a存活率的影响,应用TUNEL法和Caspase-3活性检测观察二氢杨梅素对AGZY-83-a凋亡的作用,并通过共聚焦显微镜观察二氢杨梅素对肺腺癌细胞内钙的影响。结果MTT结果显示,二氢杨梅素50μmol·L-1对AGZY-83-a的增殖有明显抑制作用,并呈剂量和时间依赖关系;TUNEL实验结果显示二氢杨梅素可以引起AGZY-83-a发生凋亡,且具有剂量依赖性;Caspase-3活性检测结果显示二氢杨梅素可增加肺腺癌细胞Caspase-3活性,说明二氢杨梅素可以诱导AGZY-83-a凋亡;二氢杨梅素可升高肺腺癌细胞内钙,平均荧光强度FI可增加20多倍。结论二氢杨梅素可通过增加细胞内钙诱导AGZY-83-a细胞凋亡,从而发挥抗肿瘤作用。

Aim To investigate the effects of dihydro- myricetin on the apoptosis in AGZY-83-a tumor cells and its relevant mechanisms. Methods The survival rate of AGZY-83-a cells was assayed by MTT dye reduction. The cellular apoptosis was analyzed by TUNEL method and caspase-3 activity. The intracellular free calcium [ Ca^2＋ ] i was detected to explore the apoptotic mechanism of dihydromyricetin. Results In the MTF assay, dihydromyricetin 50μmol · LZ^- 1 significantly inhibited survival ratios of AGZY-83-a cells at dose-and time-dependent manner. It was demonstrated in TUNEL assay that dihydromyricetin could induce the apoptosis of AGZY-83-a cells in a concentration-dependent manner. The examination of Caspase-3 activity indicated that the dihydromyricetin could induce the apoptosis of AGZY-83-a cells, which was dose-dependent activation of Caspase-3 in AGZY-83-a cells. The detection of the intracellular [ Ca^2 ＋ ] i showed that the average FI of the [ Ca^2 ＋ ] i could be markedly increased to 20-fold as the basic condition. Conclusions Di- hydromyricetin can induce the apoptosis in AGZY-83-a cells, which is associated with the overload of the in- tracellular [ Ca^2 ＋ ] i.