沉默Smad4基因对乳腺癌MCF-7细胞增殖和凋亡的影响

发布时间：2018-06-16 23:50

本文选题：Smad + MCF-细胞　；参考：《吉林大学学报(医学版)》2017年05期

【摘要】：目的:研究降低Smad4表达对乳腺癌MCF-7细胞增殖和凋亡的影响,并探讨其可能的作用机制。方法:体外培养人乳腺癌MCF-7和MDA-MB-231细胞,采用逆转录-聚合酶链反应(RT-PCR)法检测MCF-7细胞和MDA-MB-231细胞中Smad4 mRNA表达水平;Smad4-shRNA与Scramble-shRNA质粒分别稳定转染Smad4高表达MCF-7细胞,实验分为未转染MCF-7细胞组(正常对照组),采用Smad4基因沉默组和Scramble组(阴性对照组);采用CCK-8法检测各组细胞增殖能力,流式细胞术检测各组细胞凋亡率,实时荧光定量PCR(Real-time PCR)法检测增殖相关基因CDKN1A、CDK1和CDK2以及凋亡相关基因Suvivin、bcl-2、caspase 3和caspase 9mRNA表达水平。结果:各组细胞增殖能力比较差异无统计学意义(P0.05),各组细胞CDKN1A、CDK1和CDK2mRNA表达水平比较差异无统计学意义(P0.05);与正常对照组和阴性对照组比较,Smad4基因沉默组细胞凋亡率明显降低(P0.01),Smad4基因沉默组细胞Suvivin和bcl-2mRNA表达水平明显升高(P0.01),caspase 3和caspase 9mRNA表达水平明显降低(P0.05)。结论:Smad4可以通过下调Suvivin和bcl-2的表达、上调caspase 3和caspase 9表达引起细胞凋亡。
[Abstract]:Aim: to study the effect of decreasing Smad4 expression on the proliferation and apoptosis of breast cancer MCF-7 cells and to explore its possible mechanism. Methods: human breast cancer MCF-7 and MDA-MB-231 cells were cultured in vitro. The expression levels of Smad4 mRNA in MCF-7 cells and MDA-MB-231 cells were detected by reverse transcription-polymerase chain reaction (RT-PCR). Smad4-shRNA and Scramble-shRNA plasmids were stably transfected into MCF-7 cells. The experiment was divided into untransfected MCF-7 cells group (normal control group, Smad4 gene silencing group and scramble group (negative control group), CCK-8 method to detect cell proliferation ability, flow cytometry to detect cell apoptosis rate in each group, Smad4 gene silencing group and scramble group (negative control group). The expression of proliferation-related genes CDK1 and CDK2 and the apoptosis-related genes Suvivin-bcl-2 caspase-3 and caspase 9 mRNA were detected by real-time PCR. Results: there was no significant difference in cell proliferation ability among the three groups (P 0.05), but there was no significant difference in the expression of CDKN1A, CDK1 and CDK2 mRNA, and the apoptosis rate of Smad4 gene silencing group was significantly higher than that of normal control group and negative control group. The mRNA expression levels of Suvivin and bcl-2 in the P0.01Smad4 gene silencing group were significantly increased, and the expression levels of caspase-3 and caspase 9 in P0.01P0.01Smad4 gene silencing group were significantly decreased (P 0.05). Conclusion: Smad4 can up-regulate the expression of caspase _ 3 and caspase _ 9 and induce apoptosis by down-regulating the expression of Suvivin and bcl-2.
【作者单位】：北华大学基础医学院病理学教研室;吉林大学基础医学院病理生物学教育部重点实验室;北华大学化学与生物学院生命科学研究中心;吉林大学第一医院病理科;
【基金】：国家科技部支撑计划项目资助课题(2012BAI19B05)
【分类号】：R737.9

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