中国药物警戒 ›› 2016, Vol. 13 ›› Issue (10): 577-580.

• 基础与临床研究 •    下一篇

FTY720对乳腺癌细胞MCF-7的影响及机制研究

杨蕾, 吴继华, 刘佩林, 时全星, 殷召, 梁庆伟, 周丽, 董晓昕, 雷琰, 张怡堃*   

  1. 中国人民解放军第306医院,北京 100101
  • 收稿日期:2016-11-23 修回日期:2016-11-23 出版日期:2016-10-20 发布日期:2016-11-23
  • 通讯作者: 张怡堃,女,博士,副主任医师,副教授,肿瘤生物治疗研究。E-mail:yikunzhang1307@163.com
  • 作者简介:杨蕾,女,本科,副教授,临床医学。
  • 基金资助:
    国家高技术研究发展计划（863 计划）项目（2015AA80310085G）; 中国博士后基金项目(5492013M542445)

Study on Effects of FTY720 on Breast Cancer Cells MCF-7 and Its Mechanisms

YANG Lei, WU Ji-hua, LIU Pei-lin, SHI Quan-xing, YIN Zhao, LIANG Qing-wei, ZHOU Li, DONG Xiao-xi, LEI Yan, ZHANG Yi-kun*   

  1. 306th Hospital of PLA, Beijing 100101, China
  • Received:2016-11-23 Revised:2016-11-23 Online:2016-10-20 Published:2016-11-23

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摘要： 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、5 µM·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mRNA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5 μM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）。mRNA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。

关键词: FTY720, MCF-7, 凋亡, 抗肿瘤, 乳腺癌

Abstract: Objective To investigate the anti-tumor effect and preliminary mechanism of FTY720 on breast cancer cells MCF-7. Methods MCF-7 cells were incubated with different doses of FTY720 (1, 2.5, 5 µM·L-1) in vitro for 24 hours to detect the effects of FTY720 on apoptosis of MCF-7 cells by DAPI and Annexin V-APC/ PI flow cytometry. Total RNA from cells was isolated and reverse transcribed to cDNA to test the mRNA expression levels of anti-apoptotic molecule Bcl-2 and Bax by real-time PCR, further signaling pathways molecules AKT, NFκB protein expression levels were detected by Western blotting. Results FTY720 could significantly inhibit cell proliferation and induce nuclear condensation and nuclear fragmentation at a concentration of 5 µM. FTY720 could induce apoptosis rate to 30.0% at a concentration of 5 µM, and the inhibition rate could be concentration-dependent (P<0.05). The mRNA expression level of anti-apoptotic related proteins Bcl-2 was significantly down-regulated and the mRNA expression of apoptosis protein Bax was significantly up-regulated (P<0.01). We confirmed that after the MCF-7 cell being dealed with FTY720, phosphorylated AKT protein expression level was significantly reduced (P<0.05), phosphorylation NFκB protein expression level was significantly increased (P<0.05). Conclusion FTY720 can induce apoptosis in breast cancer cells by regulating the expression levels of various proteins in cells.

Key words: FTY720, MCF-7, apoptosis, antitumor, breast cancer

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