摘要
目的探讨丹参酮ⅡA（TAT）对人白血病细胞株K562的生长抑制作用及其可能机制。方法采用四甲基偶氮唑蓝（MTT）法检测TAT（终浓度1、10、15、30、50μmol／L）对K562细胞的增殖抑制作用，光学显微镜观察TAT（30μmol／L）作用24h后K562细胞的形态变化，流式细胞术检测细胞凋亡率，Westernblot法检测凋亡蛋白BCL-2、BAX的表达和蛋白激酶AKT／mTOR信号通路的变化。结果终浓度为10～50μmol／L的TAT对K562细胞的生长具有抑制作用，与对照组比较差异具有统计学意义（P〈0．05），TAT作用K562细胞96h的半数抑制浓度（IC50）值为（7．75±2．47）μmol／L。30μmol／LTAT作用K562细胞24h后显微镜下显示细胞破坏明显，部分细胞裂解成碎片。流式细胞仪检测发现30μmol／L TAT作用组凋亡细胞明显增多（P〈0．05）。Western blot结果提示，30μmol／LTAT作用K562细胞后AKT、mTOR、p70S6K、4-EBP1等信号通路关键点蛋白磷酸化表达水平减低，凋亡抑制蛋白BCL-2下调，而促凋亡蛋白BAX上调。结论TAT通过抑制AKT／m—TOR信号通路，下调BCL-2和上调BAx的表达，促进细胞凋亡，抑制白血病细胞株K562的生长。
Objective To determine the anti-proliferative effect of Tanshinone ⅡA （TAT） on leukemia K562 cell line and its mechanism. Methods The proliferation of K562 cell line was detected by MTT assay. The morphological changes of the cells were examined with light microscope. The cell apoptosis was detected by flow cytometry. The expressions of BCL-2, BAX and m-TOR signaling pathway were examined by Western blot assay. Results TAT inhibited the proliferation of K562 cell line, with a value of IC50 （7.75±2.47） μmol/L after treatment for 96 h. Significant morphological changes were found after incubation of the cells for 24 h. The apoptotic rate accelerated after TAT treatment for 24 h compared with the controls. TAT down-regulated the expression of mTOR signaling pathway and BCL-2 protein, and up-regulated proapoptotic protein BAX. Conclusion TAT can inhibit the proliferation of K562 cells through down-regulating mTOR signaling pathway, inducing apoptosis.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2014年第3期410-413,共4页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(No.30770912)
四川省科技厅社会公益项目(No.2008SZ0017)
教育部回国留学人员启动金(No.20071108-18-4)资助
