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目的探讨FTY720对乳腺癌细胞MCF-7的抗肿瘤作用并初步探讨其机制。方法体外培养乳腺癌细胞MCF-7,分别加入1、2.5、5μM·L-1的FTY720,培养24 h,观察FTY720对MCF-7细胞凋亡的影响;Q-PCR检测抗凋亡分子Bcl-2以及凋亡分子Bax的m RNA水平表达变化,Western blot检测信号通路AKT、NFκB的蛋白表达水平变化,检测FTY720对MCF-7细胞的抗凋亡作用及其作用机制。结果 FTY720浓度为5μM时MCF-7细胞增殖明显受到抑制,镜下观察可见核固缩和核碎裂,凋亡率达到30.0%,抑制率呈浓度依赖(P<0.05)。m RNA水平上,抗凋亡相关蛋白Bcl-2表达下调,凋亡蛋白Bax表达显著上调(P<0.01)。本次研究证实FTY720作用于MCF-7细胞后,蛋白磷酸化AKT表达显著下调(P<0.05),蛋白磷酸化NFκB表达显著上调(P<0.05)。结论 FTY720可诱导乳腺癌细胞MCF-7的凋亡,并且证实通过调节细胞内多种信号分子的表达水平,从而诱导细胞凋亡。
Objective To investigate the antitumor effect of FTY720 on breast cancer cell line MCF-7 and its mechanism. Methods The breast cancer cells MCF-7 were cultured in vitro. The cells were incubated with 1,2,5,5 μM · L-1 FTY720 for 24 h. The effect of FTY720 on the apoptosis of MCF-7 cells was observed. The anti-apoptotic molecules Bcl- 2 and Bax, and the protein expression levels of AKT and NFκB were detected by Western blot. The anti-apoptotic effect of FTY720 on MCF-7 cells and its mechanism were detected. Results The proliferation of MCF-7 cells was obviously inhibited when the concentration of FTY720 was 5μM. Nuclear condensation and nuclear fragmentation were found in the microscope. The apoptosis rate reached 30.0% and the inhibition rate was concentration-dependent (P <0.05). At the m RNA level, the expressions of Bcl-2 and Bax were up-regulated (P <0.01). This study confirmed that after FTY720 treatment on MCF-7 cells, protein phosphorylation of AKT was significantly down-regulated (P <0.05), and protein phosphorylation of NFκB was significantly up-regulated (P <0.05). Conclusion FTY720 can induce apoptosis of breast cancer cell line MCF-7, and it has been demonstrated that FTY720 induces apoptosis by regulating the expression of various signaling molecules in the cell.