Ge Ya-Kun, Zhang Yuan-Xin, Chen Yun-Peng, Bai Xu, Shi Wei
College of Environmental & Biological Engineering, Jilin Institute of Chemical Technology, Jilin 132022, China.
Zhonghua Zhong Liu Za Zhi. 2012 Feb;34(2):100-3. doi: 10.3760/cma.j.issn.0253-3766.2012.02.005.
To observe the effect of NL-608 (a nutlin analog) on apoptosis induction in human breast cancer MCF-7 cells in vitro, and investigate the relevant molecular mechanism.
The effect of NL-608 on proliferation of MCF-7 cells was determined by MTT assay. The apoptosis in MCF-7 cells was determined by flow cytometry with annexin V-FITC and PI. The activity of caspase 3, caspase 8 and caspase 9 was determined with caspase activity assay kit and Western blot, and the proteins of Fas and FasL were determined by Western blot.
NL-608 showed a dose-dependent inhibitory effect on the proliferation of MCF-7 cells. It induced apoptosis in MCF-7 cells in a dose-dependent manner. The activity of caspase 3 and caspase 8 in MCF-7 cells was increased with the increasing concentration of NL-608, but caspase 9 had no changes. The proteins of Fas and FasL were increased in a dose-dependent manner.
NL-608 induces apoptosis in MCF-7 cells in vitro through inducing caspase 3 activity and death receptor-mediated signal pathway.
观察NL-608（一种纽林类似物）对人乳腺癌MCF-7细胞体外诱导凋亡的作用，并探讨其相关分子机制。
采用MTT法检测NL-608对MCF-7细胞增殖的影响。采用膜联蛋白V-异硫氰酸荧光素（annexin V-FITC）和碘化丙啶（PI）流式细胞术检测MCF-7细胞凋亡情况。用半胱天冬酶（caspase）活性检测试剂盒及蛋白质免疫印迹法检测caspase 3、caspase 8和caspase 9的活性，并用蛋白质免疫印迹法检测Fas和FasL蛋白。
NL-608对MCF-7细胞的增殖呈剂量依赖性抑制作用。它以剂量依赖性方式诱导MCF-7细胞凋亡。随着NL-608浓度的增加，MCF-7细胞中caspase 3和caspase 8的活性增加，但caspase 9无变化。Fas和FasL蛋白呈剂量依赖性增加。
NL-608通过诱导caspase 3活性和死亡受体介导的信号通路在体外诱导MCF-7细胞凋亡。
