雷公藤甲素抑制未分化甲状腺癌细胞增殖

Inhibitory effects of triptolide on the proliferation of human anaplastic thyroid carcinoma FRO Cells

目的研究雷公藤甲素对人未分化甲状腺癌FRO细胞株增殖抑制作用。方法以雷公藤甲素处理FRO细胞,采用WST-1法检测细胞增殖的抑制率、Western blot方法分析雷公藤甲素对FRO细胞的activated Caspase-3和Caspase-9蛋白表达的影响;荧光探针-2’,7’-二氯氢化荧光素-二乙酸酯测定雷公藤甲素对胞内活性氧(ROS)水平的影响,酶标仪检测胞内谷胱甘肽(GSH)和氧化性谷胱甘肽(GSSG)含量的变化。结果与空白对照组相比,雷公藤甲素呈浓度、时间依赖性抑制FRO细胞增殖,IC50为22.8 nmol/L;Western blot检测发现雷公藤甲素给药处理12 h后,其activated Caspase-3和Caspase-9蛋白表达明显增加。雷公藤甲素作用2 h后,能显著增加FRO细胞ROS水平(P<0.01);雷公藤甲素能显著降低FRO细胞内GSH水平,作用48 h胞内GSH降为对照组的(21.07±1.29)%(P<0.05),而氧化型谷胱甘肽GSSG含量的变化无统计学意义(P>0.05)。结论雷公藤甲素对人未分化甲状腺癌FRO细胞株的生长有抑制作用,诱导细胞凋亡,胞内氧化还原稳态失衡在其中起了重要作用。

Objective To study the inhibitory effects of triptolide on the proliferation of human anaplastic thyroid carcinoma FRO cells. Methods FRO cells were treated in vitro with different concentration triptolide（0-100 nmol / L）for 24 h. WST-1 assay was used to detect the proliferation of FRO cells. Western blot was used to measure the expression of apoptosis related proteins activated caspase-3 and caspase-9. Intracellular reactive oxygen species（ROS）level was analyzed by H2 DCFDA fluorescence incubation and measured by spectrophotometry. Intracellular GSH and GSSG concentrations were detected. Results Compared with blank group, proliferation inhibition was increased significantly in triptolide group（P 〈0.05） in a time- and concentration-dependent manner. The levels of activated caspase-3 and caspase-9 were significantly increased after treated with triptolide for 12 h（P〈0.05）. Intracellular level of reactive oxygen species（ROS） in FRO cells was significantly increased from 2 h after treatment（P 〈0.01）. Intracellular glutathione（GSH） level was significantly reduced in FRO cells treated with triptolide for 12 h（P 〈0.05） in a concentration-dependent manner, but the GSSG level without obvious alteration（P 〉0.05）. Conclusion Triptolide could significantly inhibit the proliferation of FRO cells by up-regulating the expression of activated caspase-3 and caspase-9,and the mechanism might be due to triptolide induced redox state imbalance.