中华放射医学与防护杂志

徐晓婷,周菊英,刘蔚,等.辐射对恶性脑胶质瘤细胞放射敏感性的影响及其机制[J].中华放射医学与防护杂志,2012,32(6):602-606.XU Xiao-ting,ZHOU Ju-ying,LIU Wei,et al.Effect of radiation on the radiosensitivity of a human malignant glioma cell line[J].Chin J Radiol Med Prot,2012,32(6):602-606

辐射对恶性脑胶质瘤细胞放射敏感性的影响及其机制

Effect of radiation on the radiosensitivity of a human malignant glioma cell line

投稿时间：2012-04-05

DOI：10.3760/cma.j.issn.0254-5098.2012.06.010

中文关键词: 胶质瘤辐射抗性 cyclin B1 miR-21 Stat3

英文关键词:Glioma Radioresistance cyclin B1 miR-21 Stat3

基金项目:

作者	单位	E-mail
徐晓婷	215006 苏州大学附属第一医院肿瘤放疗科
周菊英	215006 苏州大学附属第一医院肿瘤放疗科	Zhjuying@sohu.com
刘蔚	215006 苏州大学附属第一医院肿瘤放疗科
李莉	215006 苏州大学附属第一医院肿瘤放疗科
吴琼	215006 苏州大学附属第一医院肿瘤放疗科
秦颂兵	215006 苏州大学附属第一医院肿瘤放疗科
王利利	215006 苏州大学附属第一医院肿瘤放疗科

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中文摘要:

目的观察辐射对人脑恶性胶质瘤细胞株SHG-44辐射敏感性的影响,并探讨辐射抗性产生的机制。方法使用胶质瘤细胞株SHG-44及经单次照射10 Gy后连续培养15代的辐射后细胞株SHG-44_{10 Gy},采用集落形成实验评估辐射对两种细胞株辐射敏感性的影响,流式细胞仪检测两种细胞株的辐射前后细胞周期时相分布及凋亡率,并用RT-PCR法检测cyclin B1 mRNA 和miR-21的表达水平,Western blot法检测信号转导子和转录激活子3(Stat3)蛋白表达水平。结果比较D₀、SF₂,SHG-44细胞(D₀=2.35、SF₂=0.62),低于SHG-44_{10 Gy}细胞(D₀=3.22,SF₂=0.74)。与SHG-44细胞相比,SHG-44_{10 Gy}细胞G₂/M期比例减少,S期比例增多(F=22.21,P<0.05)。照射前后SHG-44_{10 Gy} cyclinB1 mRNA 相对量大于SHG-44细胞株(t=3.1、4.1,P<0.05)。照射前后的早期凋亡率分别为SHG-44 (17.60±0.26)%和(28.00±0.36)%,SHG-44_{10 Gy}(4.20±0.30)%和(5.17±0.65)%,SHG-44_{10 Gy}细胞与SHG-44细胞相比,早期凋亡率明显下降(t=58.0,P<0.01)。qRT-PCR及Western blot法检测SHG-44_{10 Gy}细胞的miR-21表达及Stat3蛋白表达均较SHG-44升高。结论照射后SHG-44细胞辐射抗性增加,可能与辐射上调细胞周期信号传导通路中的靶基因cyclinB1导致的G₂/M期细胞比例减少有关;同时,辐射导致细胞miR-21表达增高,降低细胞的凋亡。

英文摘要:

Objective To observe the differences of the radio-biological characteristics between the human malignant glioma cell line SHG-44 and its progeny cells SHG-44_{10 Gy} and to probe the underlying mechanism. Methods The SHG-44_{10 Gy} cells were the progeny of SHG cells that had been irradiated with 10 Gy X-rays and then passaged for 15 generations. The radiosensitivity of SHG-44 and SHG-44_{10 Gy} wre measured by clonogenic assay and the multi-target single-hit model was used to fit the survival curve. The cell cycle redistribution and apoptosis were analyzed by flow cytometry assay. Quantitative Real Time-PCR (qRT-PCR) was used to determine the relative levels of cyclin B1 mRNA and miR-21. Stat3 protein levels were detected by Western blot. Results The values of D₀, D_q and SF₂ of SHG-44_{10 Gy} cells were significantly higher than those of SHG-44 cells. Flow cytometric analysis showed that there was less G₂/M phase arrest in SHG-44_{10 Gy}(F=22.21, P<0.05). Radiation-induced early apoptotic population was increased from (17.60± 0.26)% to (28.00± 0.36)% for SHG-44 cells, but increased from only (4.20± 0.30)% to (5.17± 0.65)% for SHG-44_{10 Gy}. miR-21 in SHG-44_{10 Gy} cells were increased by 1.44 fold of SHG-44 cells, which was associated with the increase of Stat3 protein expression. Conclusions Radioresistance is observed in the progeny of human malignant glioma cell line SHG-44 which had been irradiated with 10 Gy X-rays. The underlying mechanisms may be relative to the upregulation of cyclin B1 that acts as a key downstream gene in the signaling pathway of G₂/M phase transition. In addition, the upregulation of miR-21 may be involved in the apoptosis of SHG-44_{10 Gy} cells.

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