Matrine Inhibits Breast Cancer Growth Via miR-21/PTEN/Akt Pathway in MCF-7 Cells
Matrine Inhibits Breast Cancer Growth Via miR-21/PTEN/Akt Pathway in MCF-7 Cells
复制标题
苦参碱通过 MCF-7 细胞中的 miR-21/PTEN/Akt 通路抑制乳腺癌生长
DOI：
10.1159/000341444
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发表时间：
2012-01-01
影响因子：
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通讯作者：
Jiang, Hong-Chi
中科院分区：
文献类型：
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作者：
Li, Lin-Qiang;Li, Xue-Lian;Jiang, Hong-Chi
背景资料：苦参碱是从苦参中提取的主要生物碱之一，在我国临床上用于治疗乳腺癌，疗效显著。然而，其机制在很大程度上仍然未知。方法：MTT法检测细胞活力。苦参碱作用MCF-7细胞48 h后，采用流式细胞仪、TUNEL法和透射电镜检测细胞凋亡，流式细胞仪分析细胞周期分布。此外，通过Western印迹法测定PTEN、pAkt、Akt、pBad、Bad、p21/WAF 1/CIP 1和p27/KIP 1的表达。通过实时定量RT-PCR定量miR-21水平的变化。将miR-21转染MCF-7细胞后，Western blot检测PTEN蛋白表达水平。结果：苦参碱可诱导MCF-7细胞凋亡，使细胞周期阻滞于G1/S期，呈浓度和时间依赖性抑制MCF-7细胞生长。苦参碱通过下调miR-21上调PTEN，miR-21反过来使Akt去磷酸化，导致Bad、p21/WAF 1/CIP 1和p27/KIP 1的积累。结论：我们的研究首次揭示了苦参碱抑制乳腺癌生长的能力，并阐明了miR-21/PTEN/Akt通路作为苦参碱抗癌作用的信号传导机制。我们的研究结果也加强了这样一种观点，即miRNAs可以作为天然药物治疗效果的介质。
Background: Matrine is one of the major alkaloids extracted from Sophora flavescens and has been used clinically for breast cancer with notable therapeutic efficacy in China. However, the mechanisms are still largely unknown. Methods: Cell viability was analyzed by MTT assay. After MCF-7 cells were treated with matrine for 48h, apoptosis was detected by flow cytometry, TUNEL assay and transmission electron microscopy, and the cell cycle distribution was also analyzed by flow cytometry. Further, the expression of PTEN, pAkt, Akt, pBad, Bad, p21/WAF1/CIP1 , and p27/KIP1 was determined by Western blot. Changes of miR-21 level were quantified by real-time RT-PCR. After miR-21 was transfected in MCF-7 cells, PTEN protein level was measured by Western blot. Results: Matrine inhibited MCF-7 cell growth in a concentration-and time-dependent manner, by inducing apoptosis and cell cycle arrest at G1/S phase. Matrine up-regulated PTEN by downregulating miR-21 which in turn dephosphorylated Akt, resulting in accumulation of Bad, p21/WAF1/CIP1 and p27/KIP1. Conclusion: Our study unraveled, for the first time, the ability of matrine to suppress breast cancer growth and elucidated the miR-21/PTEN/Akt pathway as a signaling mechanism for the anti-cancer action of matrine. Our findings also reinforce the notion that miRNAs can act as mediators of the therapeutic efficacy of natural medicines.