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Silencing of ATM expression by siRNA technique contributes to glioma stem cell radiosensitivity in vitro and in vivo

  • Authors:
    • Yan Li
    • Luchun Li
    • Zhijuan Wu
    • Lulu Wang
    • Yongzhong Wu
    • Dairong Li
    • Uiwen Ma
    • Jianghe Shao
    • Huiqing Yu
    • Donglin Wang

  • View Affiliations

  • Published online on: May 24, 2017     https://doi.org/10.3892/or.2017.5665
  • Pages: 325-335
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Abstract

Evidence has shown that both high expression of the ataxia-telangiectasia mutated (ATM) gene and glioma stem cells (GSCs) are responsible for radioresistance in glioma. Thus, we hypothesized that brain tumor radiosensitivity may be enhanced via silencing of the ATM gene in GSCs. In the present study we successfully induced GSCs from two cell lines and used CD133 and nestin to identify GSCs. A lentivirus was used to deliver siRNA-ATMPuro (A group) to GSCs prior to radiation, while siRNA-HKPuro (N group) and GSCs (C group) were used as negative and blank controls, respectively. RT-qPCR and western blotting were performed to verify the efficiency of the siRNA-ATM technique. The expression of the ATM gene and ATM protein were significantly downregulated post-transfection. Cell Counting Kit-8 (CCK-8) and colony formation assays revealed that the A group demonstrated weak cell proliferation and lower survival fractions post-irradiation compared to the C/N groups. Flow cytometry was used to examine the percentage of cell apoptosis and G2 phase arrest, which were both higher in the A group than in the C/N groups. We found that the comet tail percentage evaluated by comet assay was higher in the A group than in the C/N groups. After radiation treatment, three radiosensitive genes [p53, proliferating cell nuclear antigen (PCNA), survivin] exhibited a decreasing tendency as determined by RT-qPCR. Mice underwent subcutaneous implantation, followed by radiation, and the resulting necrosis and hemorrhage were more obvious in the A group than in the N groups. In conclusion, silencing of ATM via the siRNA technique improved radiosensitivity of GSCs both in vitro and in vivo.
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July-2017
Volume 38 Issue 1

Print ISSN: 1021-335X
Online ISSN:1791-2431

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  • Source Normalized Impact per Paper (SNIP): 0.786
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Spandidos Publications style
Li Y, Li L, Wu Z, Wang L, Wu Y, Li D, Ma U, Shao J, Yu H, Wang D, Wang D, et al: Silencing of ATM expression by siRNA technique contributes to glioma stem cell radiosensitivity in vitro and in vivo. Oncol Rep 38: 325-335, 2017
APA
Li, Y., Li, L., Wu, Z., Wang, L., Wu, Y., Li, D. ... Wang, D. (2017). Silencing of ATM expression by siRNA technique contributes to glioma stem cell radiosensitivity in vitro and in vivo. Oncology Reports, 38, 325-335. https://doi.org/10.3892/or.2017.5665
MLA
Li, Y., Li, L., Wu, Z., Wang, L., Wu, Y., Li, D., Ma, U., Shao, J., Yu, H., Wang, D."Silencing of ATM expression by siRNA technique contributes to glioma stem cell radiosensitivity in vitro and in vivo". Oncology Reports 38.1 (2017): 325-335.
Chicago
Li, Y., Li, L., Wu, Z., Wang, L., Wu, Y., Li, D., Ma, U., Shao, J., Yu, H., Wang, D."Silencing of ATM expression by siRNA technique contributes to glioma stem cell radiosensitivity in vitro and in vivo". Oncology Reports 38, no. 1 (2017): 325-335. https://doi.org/10.3892/or.2017.5665