小干扰RNA下调Notch-1基因的表达对三阴性乳腺癌
MDA-MB-231细胞生物学功能的影响
潘红;凌立君;周文斌;梁秀清;王水
【期刊名称】《中华实验外科杂志》 【年(卷),期】2012(029)005
【摘要】目的 观察Notch-1信号通路在三阴性乳腺癌(TNBC)细胞MDA-MB-231增殖中的作用,并探讨其分子机制.方法 运用小干扰RNA(siRNA)转染技术,下调MDA-MB-231细胞中Notch-1 mRNA表达,应用噻唑蓝(MTT)比色法检测干扰后5d的细胞增殖,并用流式细胞术检测干扰48 h后细胞增殖、凋亡及细胞周期的变化.另外应用实时定量聚合酶链反应(PCR)和Western blot法检测siRNA转染前后Notch-1、细胞周期素D1( Cyclin D1)、B细胞淋巴瘤/白血病-2基因(bcl-2)、bcl-XL mRNA变化及细胞核中核因子(NF)-κB蛋白的变化.结果 N0tch-1 siRNA可有效降低细胞中Notch-1 mRNA表达；转染后细胞增殖能力明显受到抑制,第5天的抑制率达到44.7％,细胞凋亡率由3.67％增加到13.25％,细胞周期G2期由6.27％增加到14.28％.进一步研究结果显示siRNA介导的Notch-1下调,可使细胞核内NF-κB蛋白表达降低,细胞中Cyclin D1、bcl-2、bcl-XL mRNA 表达下调.结论 Notch-1信号通路在TNBC中发挥重要作用,siRNA下调Notch-1可有效抑制MDA-MB-231细胞增殖,Notch-1是TNBC潜在的新治疗靶点.%Objective To investigate the role of Notch-1 signaling pathway in the growth of triplenegative breast cancer ( TNBC ) and the action mechanism.Methods Notch-1 small interfering RNA (siRNA) was transiently transfected into MDA-MB-231 cells to down-
regulate the mRNA expression of Notch-1.The cell proliferation was assessed by methyl thiazol tetrazolium (MTT) assay during 5 days after transfection.And the changes in cell apoptosis and cell cycle were evaluated 48 h after transfection by flow cytometry.Real-time polymerase chain reaction (PCR) was used to detect the mRNA expression of Notch-1,Cyclin D1,bcl-2,bcl-XL and Western blotting was used to measure nuclear factor-κB (NF-κB) level in cell nucleus.Results Notch-1 siRNA could effectively inhibit the expression of Notch-1,with the inhibition rate being 88.6％.The proliferation of MDA-MB-231 cells was markedly inhibited by Notch-1 siRNA with the inhibition rate being 44.7％.The apoptosis rate was increased from 3.67％ to 13.25％ and the percentage of cells in G2 phase was increased from 6.27％ to 14.28％,respectively.Down-regulation of the expression of Notch-1 by siRNA could down-regulate the expression of NF-κB in nucleus,and the expression
of
Cyclin
D1,bcl-2
and
bcl-XL
in
MDA-MB-231
cells.Conclusion Our results suggest that Notch-1 pathway plays a critical role in TNBC.Down-regulation of Notch-1 by siRNA can inhibit the proliferation of MDA-MB-231 cells.Notch-1 may be a novel therapeutic target of TNBC. 【总页数】4页(797-800)
【关键词】三阴性乳腺癌;Notch-1;核因子-κB;RNA干扰 【作者】潘红;凌立君;周文斌;梁秀清;王水
【作者单位】210029 南京医科大学第一附属医院普外科;210029 南京医科大学第一附属医院普外科;210029 南京医科大学第一附属医院普外科;210029 南京医科大学第一附属医院普外科;210029 南京医科大学第一附属医院普外科 【正文语种】中文 【中图分类】 【文献来源】
https://www.zhangqiaokeyan.com/academic-journal-cn_chinese-journal-experimental-surgery_thesis/0201234794555.html 【相关文献】
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小干扰RNA下调Notch-1基因的表达对三阴性乳腺癌MDA-MB-231细胞生物学功
