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靶向notch1的siRNA对U87-EGFRvⅢ胶质瘤细胞株凋亡和放射敏感度的影响[J]. 肿瘤防治研究, 2016, 43(4): 245-248. DOI: 10.3971/j.issn.1000-8578.2016.04.001
引用本文: 靶向notch1的siRNA对U87-EGFRvⅢ胶质瘤细胞株凋亡和放射敏感度的影响[J]. 肿瘤防治研究, 2016, 43(4): 245-248. DOI: 10.3971/j.issn.1000-8578.2016.04.001
Effect of siRNA-mediated notch1 Gene Silencing on Apoptosis and Radiosensitivity of Glioma Cell Line U87-EGFRvⅢ[J]. Cancer Research on Prevention and Treatment, 2016, 43(4): 245-248. DOI: 10.3971/j.issn.1000-8578.2016.04.001
Citation: Effect of siRNA-mediated notch1 Gene Silencing on Apoptosis and Radiosensitivity of Glioma Cell Line U87-EGFRvⅢ[J]. Cancer Research on Prevention and Treatment, 2016, 43(4): 245-248. DOI: 10.3971/j.issn.1000-8578.2016.04.001

靶向notch1的siRNA对U87-EGFRvⅢ胶质瘤细胞株凋亡和放射敏感度的影响

Effect of siRNA-mediated notch1 Gene Silencing on Apoptosis and Radiosensitivity of Glioma Cell Line U87-EGFRvⅢ

  • 摘要: 目的 探讨靶向notch1的siRNA对过表达EGFRvⅢ的U87胶质瘤细胞株U87-EGFRvⅢ凋亡和放射敏感度的影响。方法 利用脂质体转染合成的siRNA转染U87-EGFRvⅢ细胞。用RT-PCR和免疫印迹法来分别检测 notch1的蛋白和mRNA的表达水平;用AnnexinⅤ-FITC/PI流式细胞术检测细胞凋亡情况;用平板集落形成实验检测转染后细胞的放射敏感度。结果 合成的notch1 siRNA可有效抑制notch1在mRNA和蛋白水平的表达(P<0.05)。转染notch1 siRNA作用后细胞凋亡率比对照组明显增加(P<0.05)。平板集落形成实验初步证明notch1 siRNA可提高U87-EGFRvⅢ细胞的放射敏感度。结论 下调notch1基因可促进U87-EGFRvⅢ细胞的凋亡和提高其放射敏感度,为研究notch1基因在肿瘤放疗中的作用及其靶向联合治疗胶质瘤提供基础。

     

    Abstract: Objective To investigate the effect of notch1 siRNA blockade on the apoptosis and radiosensitivity of human glioma cell line U87-EGFRvⅢ. Methods All siRNAs were transfected into U87-EGFRvⅢ cells by Lipofectamine. The protein expression and mRNA levels of notch1 gene were detected by Western blot and RT-PCR. The AnnexinⅤ-FITC/PI assays were used to detect the inhibitory effect on cell apoptosis. The radiosensitivity parameters of U87-EGFRvⅢ cells were obtained by colony forming assay. Results The expression of notch1 mRNA and protein were significantly reduced by notch1 siRNA(P<0.05). The notch1 siRNA could significantly increase the cell apoptotic rates, compared with the control group(P<0.05). The primary result of colony forming assay showed that notch1 siRNA could enhance the radiosensitivity of U87-EGFRvⅢ cells. Conclusion Down-regulating notch1 expression could enhance the apoptosis and radiosensitivity of U87-EGFRvⅢ cells, which provides useful information for the basic research of notch1 gene in the radiotherapy and combined targeted therapy on glioma.

     

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