Department of Neurology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Department of Radiology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Cancer Med. 2019 Mar;8(3):1175-1185. doi: 10.1002/cam4.1971. Epub 2019 Feb 10.
We focused on the effects of PTGS2/NF-κB signaling pathway on the radiation resistance of glioma in the study.
We downloaded the microarray data from the Gene Expression Omnibus (GEO) database. We verified transfection successfully through QRT-PCR analysis. Immunofluorescence was used to detect γH2AX content under 2 Gy radiation. The survival rates of cells under 2 Gy irradiation were tested by clonogenic survival assay. Flow cytometry was used to detect cell cycle. Western blot was applied to detect the expression of NF-κB pathway-related proteins. We also used MTT assay to detect the proliferation of cells.
In this research, we discovered that the expression of the PTGS2 was upregulated in radiation-resistant glioma cells. The radio-tolerance rate of U87 cells was obviously elevated after the overexpression of PTGS2. The radioresistance of U87R cells was significantly reduced after the knockdown of PTGS2. After radiotherapy, the number of cells arrested in G2/M phase decreased after PTGS2 overexpression in U87cells but increased in PTGS2 knockdown in U87R cells. The survival rate of U87 and U87R cells under radiation decreased significantly after the addition of NF-κB inhibitor. The proliferation of U87 cells was suppressed by radiation and the addition of Bay 11. In addition, PTGS2 activated NF-κB signaling pathway and prevented DNA damage after radiotherapy. Lastly, PTGS2 was proved to facilitate tumor cell proliferation and improve the radio-tolerance.
PTGS2/NF-κB signaling pathway was involved in radio-tolerance of glioma cells, which provided a new insight into glioma therapy.
本研究聚焦于 PTGS2/NF-κB 信号通路对脑胶质瘤放射抵抗的影响。
从基因表达综合数据库(GEO)中下载微阵列数据。通过 QRT-PCR 分析验证转染是否成功。免疫荧光法检测 2Gy 辐射下 γH2AX 含量。克隆形成实验检测 2Gy 照射下细胞的存活率。流式细胞术检测细胞周期。Western blot 检测 NF-κB 通路相关蛋白的表达。MTT 检测细胞增殖。
在本研究中,我们发现 PTGS2 在放射抵抗脑胶质瘤细胞中表达上调。PTGS2 过表达后 U87 细胞的耐辐射率明显升高。PTGS2 敲低后 U87R 细胞的放射抵抗性明显降低。放射治疗后,U87 细胞中 PTGS2 过表达后 G2/M 期阻滞的细胞数量减少,但 U87R 细胞中 PTGS2 敲低后增加。U87 和 U87R 细胞在辐射下的存活率明显降低,加入 NF-κB 抑制剂后。辐射抑制 U87 细胞的增殖,加入 Bay 11 后。此外,PTGS2 激活 NF-κB 信号通路并防止放射后 DNA 损伤。最后,PTGS2 被证明促进肿瘤细胞增殖并提高放射耐受力。
PTGS2/NF-κB 信号通路参与脑胶质瘤细胞的放射耐受,为胶质瘤治疗提供了新的思路。