| 王鹏飞,黎文汉,洪顺明,张春智.miR-221/222激活Akt通路增加恶性胶质瘤的放射抗性[J].中华放射医学与防护杂志,2015,35(3):177-182 |
| miR-221/222激活Akt通路增加恶性胶质瘤的放射抗性 |
| miR-221/222 enhances radiation resistance of glioblastoma by Akt pathway |
| 投稿时间：2014-03-29 |
| DOI：10.3760/cma.j.issn.0254-5098.2015.03.004 |
| 中文关键词: 恶性胶质瘤 放射抗性 MiR-221/222 DNA损伤 Akt |
| 英文关键词:Glioblastoma Radiation resistance MiR-221/222 DNA damage Akt |
| 基金项目:天津市自然科学基金 (13JCYBJC21700);天津市卫生局公关项目(12KG113) |
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| 中文摘要: |
| 目的 探讨miR-221/222增加恶性胶质瘤放射抗性的机制。方法 实时荧光定量PCR(real-time PCR)技术检测U251、U87及LN229系胶质瘤细胞在接受照射后3和6 h miR-221/222的表达水平;通过平板克隆形成实验,观察敲低U251、U87及LN229系胶质瘤细胞miR-221/222后肿瘤细胞放射敏感性的变化;染色质免疫沉淀技术(ChIP)检测c-jun与miR-221/222的结合情况;虫荧光素酶活性报告试验验证PTEN是miR-221/222的靶基因;使用Western blot检测敲低胶质瘤细胞miR-221/222后的相关蛋白的表达;通过胶质瘤裸鼠模型,观察敲低miR-221/222后放射治疗的效果。结果 U251、U87及LN229系胶质瘤细胞在接受照射后miR-221/222的表达上调(t=5.48~29.21,P<0.05);敲低胶质瘤细胞miR-221/222后,其放射敏感性提高(F=1 202.22,1 789.12,1 012.32,P<0.05); c-jun转录调控miR-221/222的表达; PTEN是miR-221/222的靶基因(t=13.16,P<0.05);Western blot检测显示,敲低胶质瘤细胞miR-221/222后,pAkt及DNA-PKcs的表达下调,PTEN和GSK-3β的表达上调,Akt表达保持稳定;体内实验结果显示,敲低miR-221/222的表达联合放射治疗可显著抑制胶质瘤的生长(F=56.36,P<0.05)。结论 放射可诱导胶质瘤细胞miR-221/222的表达上调,miR-221/222通过激活Akt通路增加恶性胶质瘤的放射抗性。 |
| 英文摘要: |
| Objective To study the pathway of miR-221/222 in enhancing radiation resistance of glioblastoma.Methods After 2 Gy of X-ray irradiation, the expressions of miR-221/222 in U251,U87 and LN229 glioblastma cells were detected with real-time PCR. Clonogenic assay was used to measure the radiosensitivity of glioblastoma after knocking down miR-221/222. ChIP assay was used to identify the combination situation of c-jun and miR-221/222. Luciferase assay was applied to check whether PTEN was a target of miR-221/222. Western blot was used to detect the expression of relevant proteins in the glioblastoma cells after knocking down miR-221/222. The effect of miR-221/222 and irradiation on growth of glioblastoma in nude mice was also observed.Results The expression of miR-221/222 was increased by irradiation(t=5.48~29.21,P<0.05) and the radiosensitivity of anti-miR-221/222-transfected cells was also increased(F=1 202.22,1 789.12,1 012.32,P<0.05). MiR-221/222 was transcriptionally regulated by c-jun with a target of PTEN(t=13.16,P<0.05). When miR-221/222 was knocked-down, the expression of pAkt and DNA-PKcs were down-regulated while PTEN and GSK-3β were up-regulated, and the expression of Akt were not changed. Moreover, the growth of xenograft tumor was significantly inhibited by the combination treatment of anti-miR-221/222 and irradiation(F=56.36,P<0.05).Conclusions The expression of miR-221/222 in glioblastoma cells can be increased by irradiation, and the activation of Akt pathway downstream miR-221/222 could enhance the radiation resistance of glioblastoma. |
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