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沉默Toll样受体4表达对肺腺癌A549细胞凋亡及细胞周期的影响

郭隽馥, 王艳杰, 苗兰英, 丛培玮

郭隽馥, 王艳杰, 苗兰英, 丛培玮. 沉默Toll样受体4表达对肺腺癌A549细胞凋亡及细胞周期的影响[J]. 肿瘤防治研究, 2015, 42(03): 225-228. DOI: 10.3971/j.issn.1000-8578.2015.03.003
引用本文: 郭隽馥, 王艳杰, 苗兰英, 丛培玮. 沉默Toll样受体4表达对肺腺癌A549细胞凋亡及细胞周期的影响[J]. 肿瘤防治研究, 2015, 42(03): 225-228. DOI: 10.3971/j.issn.1000-8578.2015.03.003
GUO Junfu, WANG Yanjie, MIAO Lanying, CONG Peiwei. Effect of Silencing Toll-like Receptor 4 Expression on Cell Cycle and Apoptosis of Human Lung Adenocarcinoma Cells A549[J]. Cancer Research on Prevention and Treatment, 2015, 42(03): 225-228. DOI: 10.3971/j.issn.1000-8578.2015.03.003
Citation: GUO Junfu, WANG Yanjie, MIAO Lanying, CONG Peiwei. Effect of Silencing Toll-like Receptor 4 Expression on Cell Cycle and Apoptosis of Human Lung Adenocarcinoma Cells A549[J]. Cancer Research on Prevention and Treatment, 2015, 42(03): 225-228. DOI: 10.3971/j.issn.1000-8578.2015.03.003

沉默Toll样受体4表达对肺腺癌A549细胞凋亡及细胞周期的影响

基金项目: 国家自然科学基金(81202789);辽宁省教育厅高等学校科研项目(2009A500);辽宁省博士启动基金项目(20111134)
详细信息
    作者简介:

    郭隽馥(1983-),女,硕士,实验师,主要从事肿瘤分子遗传学方面的基础研究

    通讯作者:

    王艳杰,E-mail:15940157054@163.com

  • 中图分类号: R730.2;R734.2

Effect of Silencing Toll-like Receptor 4 Expression on Cell Cycle and Apoptosis of Human Lung Adenocarcinoma Cells A549

  • 摘要: 目的 探讨Toll样受体4(Toll-like receptor 4,TLR4)对肺腺癌A549细胞凋亡及细胞周期的影响。方法 体外化学合成针对TLR4的小干扰RNA(TLR4-siRNA),通过脂质体介导转染人肺腺癌A549细胞。采用Real-time PCR和流式细胞仪检测分析干扰后A549细胞中TLR4 mRNA及蛋白的表达情况。另外,采用流式细胞仪检测分析干扰72 h后细胞凋亡率和细胞周期的变化。结果 与对照组比较,转染TLR4-siRNA后,A549细胞中TLR4 mRNA及蛋白的表达显著下降(P<0.05),细胞的凋亡率明显增加[(1.73 ± 0.32)% vs.(7.40 ± 0.75)%(P<0.01)],细胞周期出现G0/G1期停滞[(48.21 ± 1.15)%vs.(66.26 ± 2.45)%(P<0.05)],同时S期细胞比例明显减少[(34.25 ± 1.46)% vs.(22.63 ± 3.39)%(P<0.05)]。结论 TLR4-siRNA能有效沉默A549细胞中TLR4的表达,在促进细胞凋亡的同时能够影响细胞周期分布从而抑制肿瘤细胞的生长,特异性干预TLR4基因的表达有望成为治疗肺癌的一种新手段。

     

    Abstract: Objective To explore the effect of Toll-like receptor 4 (TLR4) on cell cycle and apoptosis of human lung adenocarcinoma cells A549. Methods TLR4 specific small interfering RNA (TLR4-siRNA) was synthesized in vitro and transfected into A549 cells by Lipofectamine. The expression level of TLR4 mRNA and protein were examined by Real-time PCR and FCM. In addition, the apoptosis ratio and cell cycle were examined by FCM 72h after transfection. Results Compared with control group, TLR4-siRNA could significantly inhibit the expression of TLR4 mRNA and protein(P<0.05); the apoptosis ratio was significantly increased [(1.73 ± 0.32)% . (7.40 ± 0.75)% (P<0.01)], cell cycle was arrested in G0/G1 phase [(48.21 ± 1.15)% vs. (66.26 ± 2.45)% (P<0.05)] and the percentage of S phase cells were decreased significantly [(34.25 ±1.46)% vs. (22.63 ± 3.39)% (P<0.05)]. Conclusion TLR4-siRNA could effectively silence TLR4 expression in A549 cells, promote cell apoptosis and affect cell cycle to inhibit the growth of tumor cells. The specific intervention of TLR4 gene expression may be a new method for therapy of lung cancer.

     

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出版历程
  • 收稿日期:  2014-09-02
  • 修回日期:  2014-11-10
  • 刊出日期:  2015-03-24

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