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RNA干扰沉默MSI1表达对人脑胶质瘤细胞化疗敏感度的影响

刘细国, 付锴, 江普查, 宫睿

刘细国, 付锴, 江普查, 宫睿. RNA干扰沉默MSI1表达对人脑胶质瘤细胞化疗敏感度的影响[J]. 肿瘤防治研究, 2016, 43(9): 754-757. DOI: 10.3971/j.issn.1000-8578.2016.09.005
引用本文: 刘细国, 付锴, 江普查, 宫睿. RNA干扰沉默MSI1表达对人脑胶质瘤细胞化疗敏感度的影响[J]. 肿瘤防治研究, 2016, 43(9): 754-757. DOI: 10.3971/j.issn.1000-8578.2016.09.005
LIU Xiguo, FU Kai, JIANG Pucha, GONG Rui. Effect of RNA Interference Decreasing MSI1 Expression on Chemosensitivity of Human Glioma Cells[J]. Cancer Research on Prevention and Treatment, 2016, 43(9): 754-757. DOI: 10.3971/j.issn.1000-8578.2016.09.005
Citation: LIU Xiguo, FU Kai, JIANG Pucha, GONG Rui. Effect of RNA Interference Decreasing MSI1 Expression on Chemosensitivity of Human Glioma Cells[J]. Cancer Research on Prevention and Treatment, 2016, 43(9): 754-757. DOI: 10.3971/j.issn.1000-8578.2016.09.005

RNA干扰沉默MSI1表达对人脑胶质瘤细胞化疗敏感度的影响

详细信息
    作者简介:

    刘细国(1980-),男,博士,主治医师,主要从事脑肿瘤的基础与临床研究

    通讯作者:

    付锴.E-mail: kaizige@hotmail.com

  • 中图分类号: R739.41

Effect of RNA Interference Decreasing MSI1 Expression on Chemosensitivity of Human Glioma Cells

More Information
  • 摘要:
    目的 

    探讨RNA干扰沉默MSI1表达对人脑胶质瘤细胞化疗药物敏感度的影响。

    方法 

    构建针对MSI1 mRNA的小分子RNA干扰(siRNA)重组质粒pSIREN-MSI1,转染人脑胶质瘤U-87MG细胞;Western blot检测U-87MG细胞内MSI1蛋白的表达;CCK-8法观察U-87MG细胞对化疗药物替莫唑胺(TMZ)敏感度的改变;流式细胞仪(FCM)检测pSIREN-MSI1转染后对U-87MG细胞凋亡的影响。

    结果 

    成功构建了重组质粒pSIREN-MSI1,并成功转染U-87MG细胞;Western blot结果显示pSIREN-MSI1抑制U-87MG细胞中MSI1蛋白的表达(P<0.01);CCK-8结果显示在替莫唑胺作用下,pSIREN-MSI1组U-87MG细胞的存活率明显下降(P<0.01);FCM结果表明pSIREN-MSI1转染后明显提高U-87MG细胞的凋亡率(P<0.01)。

    结论 

    MSI1 siRNA能特异性抑制人脑胶质瘤细胞U-87MG细胞中MSI1的表达,增强人脑胶质瘤U-87MG细胞对化疗药物的敏感度,并促进细胞的凋亡。

     

    Abstract:
    Objective 

    To explore the effect of RNA interference decreasing MSI1 gene expression on the chemosensitivity of human glioma cells U-87MG.

    Methods 

    The design and synthesis of Msi1-specific siRNA and the transfection of U-87MG cells were conducted. The expression of MSI1 in U-87MG cells was detected by Western blot. CCK-8 was employed to observe the sensitivity of U-87MG cells to temozolomide(TMZ). FCM was used to observe the apoptosis rate of U-87MG cells after pSIREN-MSI1 transfection.

    Results 

    Lentiviral vector-mediated MSI1-siRNA was successfully constructed. Immunofluorescence assay demonstrated that the transfection efficiency was above 60%. Western blot analyses demonstrated that pSIREN-MSI1 could significantly inhibit the expression of MSI1 in U-87MG cells (P<0.01); CCK-8 results showed that when U-87MG cells were exposed to temozolomide, the growth inhibiting rate of pSIREN-MSI1 transfected cells was significantly increased, compared with those of blank control group and negative control group (P<0.01). FCM results showed that the apoptosis rate of U-87MG cells were increased after pSIREN-MSI1 transfection(P<0.01).

    Conclusion 

    MSI1-siRNA could significantly inhibit the expression of MSI1 in human glioma cells U-87MG. It could induce the apoptosis and enhance the chemosensitivity of U-87MG cells.

     

  • 图  1   Western blot检测pSIREN- MSI1转染U-87MG细胞后MSI1蛋白的表达

    Figure  1   Expression of MSI1 protein in U-87MG cells after pSIREN-MSI1 transfection detected by Western blot

    图  2   CCK-8法检测不同浓度替莫唑胺作用于各组转染细胞后的细胞存活率

    Figure  2   Survival rates of U-87MG cells treated with TMZ in each group detected by CCK-8

    图  3   流式细胞仪检测pSIREN-MSI1转染U-87MG细胞后的细胞凋亡率

    Figure  3   Apoptosis rate of U-87MG cells after pSIREN-MSI1 transfection detected by FCM

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出版历程
  • 收稿日期:  2015-08-04
  • 修回日期:  2015-11-26
  • 网络出版日期:  2024-02-04
  • 刊出日期:  2016-08-31

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