miR-34a通过靶向调控Notch1表达影响乳腺癌细胞的增殖
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Inhibitory effect of miR-34a on the proliferation of breast cancer cell by targeting Notch1
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    摘要:

    目的 探讨miR-34a通过靶向调控Notch1表达影响乳腺癌细胞的增殖。方法 乳腺癌细胞MCF-7转染miR-34a mimics及miR-34a NC,RT-PCR法检测细胞中miR-34a表达,MTT,克隆形成实验,Hoechst染色及流式细胞术分别检测miR-34a mimics对乳腺癌MCF-7细胞活力、克隆能力、凋亡情况及细胞周期的影响;通过双荧光素酶报告基因实验,western blot及RT-PCR检测Notch1是否是miR-34a的下游靶基因。结果 与miR-34a NC比较,miR-34 mimics组中miR-34a表达量上调,细胞活力降低(P<0.01),细胞凋亡率显著提高(P<0.01),细胞克隆数目减少(P<0.01),细胞周期阻滞在G1期(P<0.01),Notch1蛋白及mRNA表达量都显著降低(P<0.01),同时荧光素酶报告基因实验也证实miR-34a mimics能与报告基因结合。结论 miR-34a mimics能通过负性调控Notch1表达,从而显著的抑制MCF-7乳腺癌细胞增值,并诱导细胞凋亡。

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    Objective To explore inhibitory effect of miR-34a on the proliferation of breast cancer cell MCF-7 by targeting Notch1. Methods MCF-7 cells were transferred with miR-34a mimics and miR-34a NC. The expression of miR-34a in MCF-7 cell was examed by RT-PCR. The cell viability was detected by MTT. The cell apoptosis was examed by Hoechst staining. Cell cycle was detected by flow cytometry. Western blot, RT-PCR and dual luciferase reporter gene assay was to detect whether Notch1 was a downstream target gene of miR-34a. Results Compared with miR-34a NC group, the expression of miR-34a was up-regulated, the cell viability was decreased, cell apoptotic rate was increased, cell cycle was arrested in the G1 phase, the expression of Notch1 protein and mRNA was down-regulated (P<0.01). Conclusion miR-34a mimics inhibited MCF-7 cell proliferation and induced cell apoptosis via targeting regulation expression of Notch1.

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高砚春. miR-34a通过靶向调控Notch1表达影响乳腺癌细胞的增殖[J].中国比较医学杂志,,().

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  • 收稿日期:2016-06-30
  • 最后修改日期:2016-07-18
  • 录用日期:2016-07-18
  • 在线发布日期: 2016-11-28
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