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中国癌症防治杂志

中国癌症防治杂志 ›› 2020, Vol. 12 ›› Issue (2): 162-168.doi: 10.3969/j.issn.1674-5671.2020.02.10

• 基础研究 • 上一篇    下一篇

盐霉素通过靶向调控ALDH影响三阴性乳腺癌细胞MDA-MB- 231增殖和凋亡

  

  1. 延安大学医学院；延安市肿瘤防治研究重点研究室
  • 出版日期:2020-04-25 发布日期:2020-05-05
  • 通讯作者: 赵菊梅 E-mail：jmz2003.stu@163.com
  • 基金资助:
    国家自然科学基金项目（81470140); 陕西省教育厅2019专项科学研究计划项目（19JK0970）

Effect of salinomycin on the proliferation and apoptosis of triple negative breast cancer cell line MDA-MB-231 by target-regulating ALDH

  1. Medical College of Yan'an University；Key Laboratory of the Prevention and Treatment of Tumor
  • Online:2020-04-25 Published:2020-05-05

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摘要： 目的 探讨盐霉素（salinomycin，Sal）对三阴性乳腺癌细胞MDA-MB-231增殖和凋亡的影响及其作用机制。方法 采用流式细胞术筛选ALDH高表达乳腺癌细胞系；采用MTT法检测不同浓度Sal（1~32 μmol/L）对细胞增殖的影响；采用流式细胞仪检测Sal对细胞凋亡以及肿瘤干细胞表面标志物ALDH表达的影响；采用qRT-PCR、Western blot检测凋亡相关分子Caspase-3、Bax、Bcl-2、PARP mRNA和蛋白表达；TCGA（the Cancer Genome Atlas）数据库下载乳腺癌患者数据集（n=1 218），采用Pearson法分析ALDH与凋亡相关分子表达的相关性。结果 Sal在24 h、48 h、72 h均可以剂量依赖的方式抑制MDA-MB-231细胞增殖（均P<0.001）；Sal作用MDA-MB-231细胞48 h后，与对照组比较，2 μmol/L组、4 μmol/L组、8 μmol/L组的细胞凋亡率均升高（均P<0.05）；Bcl-2、PARP、Caspase-3 mRNA表达下降（均P<0.05），Bax mRNA表达升高（均P<0.05）；Bcl-2、Caspase-3 蛋白表达下降（均P<0.05），PARP、Bax 蛋白表达升高（均P<0.05）；MDA-MB-231乳腺癌干细胞表面标志物ALDH表达下降（均P<0.05）。TCGA数据库分析显示，ALDH与抗凋亡分子Bcl-2、Caspase-3的表达呈正相关（r=0.209，P=0.007；r=0.235，P=0.002），与Bax、PARP的表达呈负相关（r=-0.326，P＜0.001；r=-0.453，P＜0.001）。结论 Sal可抑制人乳腺癌MDA-MB-231细胞生长，可能通过下调肿瘤干细胞表面标志物ALDH表达实现。

关键词: 乳腺癌, 盐霉素, 增殖, 凋亡, ALDH

Abstract: Objective To investigate the effect of salinomycin（Sal） on the proliferation and apoptosis of triple negative breast cancer cell line MDA-MB-231 and its mechanism. Methods The flow cytometry was used to screen ALDH highly expressed breast cancer cell lines，and MTT method was used to detect the effect of different concentrations of Sal（1-32 μmol/L） on cell proliferation. The flow cytometry was used to detect the effect of Sal on apoptosis and the expression of ALDH on tumor stem cell surface，qPCR and Western blot were used to detect the mRNA and protein expression of apoptosis-related molecules Caspase-3，Bax，Bcl-2 and PARP. The data of breast cancer patients were downloaded from the Cancer Genome Atlas （TCGA） database（n=1 218），and Pearson method was used to analyze the correlation between ALDH and apoptosis-related molecules expression. Results Sal could inhibit the proliferation of MDA-MB-231 cells in a dose-dependent manner at 24 h，48 h，and 72 h（all P<0.001）. Compared with the control group，the apoptosis rates of the 2 μmol/L group，4 μmol/L group and 8 μmol/L group increased after Sal acting on MDA-MB-231 cells for 48 h（all P<0.05），and the mRNA expression of Bcl-2，PARP and Caspase-3 decreased （all P<0.05），while the mRNA expression of Bax increased（all P<0.05）；the protein expression of Bcl-2 and Caspase-3 decreased（all P<0.05），while the protein expression of PARP and Bax increased（all P<0.05）； the expression of ALDH，a surface marker of tumor stem cells decreased（P<0.05）. TCGA database analysis showed that ALDH was positively correlated with the expression of anti-apoptotic molecules Bcl-2 and Caspase-3（r=0.209，P=0.007；r=0.235，P=0.002），whereas negatively correlated with the expressions of Bax and PARP（r=-0.326，P<0.001； r=-0.453，P<0.001）. Conclusion Sal can inhibit the growth of human breast cancer cell line MDA-MB-231，possibly by down-regulating the expression of ALDH，a surface marker of tumor stem cells.

Key words: Breast cancer, Salinomycin, Proliferation, Apoptosis, ALDH

中图分类号: 

  • R737.9