摘要
目的：通过检测放射线对胶质瘤U251细胞β-catenin与去泛素化酶USP9X表达水平的影响,以期了解与胶质瘤细胞放射后去泛素化酶USP9X及Wnt信号通路重要因子β-catenin表达的变化及其意义。方法：胶质瘤细胞U251细胞经过不同放疗剂量照射后,Real time PCR法检测细胞中USP9X与β-catenin基因表达,Western Blotting法检测USP9X与β-catenin的蛋白表达,流式细胞术检测不同放射剂量处理后U251细胞凋亡水平。
结果：Real Time PCR结果显示在1Gy放疗剂量照射时USP9X和β-catenin的表达水平升高,随后随放疗剂量增加,USP9X和β-catenin基因表达成下降趋势,Western Blotting结果显示USP9X、β-catenin的蛋白表达量与其基因表达水平成正相关,凋亡实验结果显示细胞凋亡率与放射剂量成正相关。
结论：在1Gy低剂量照射时USP9X和β-catenin的表达水平最高,证明小剂量放疗可能提高U251细胞的放射抵抗,增加DNA修复。随后随放疗剂量增加,USP9X和β-catenin的表达成下降趋势,,说明大剂量放疗使U251细胞不足以完成细胞DNA修复,细胞凋亡增加。证明USP9X与胶质瘤细胞的放射后凋亡水平相关。
Objective To research on the relationship between deubiquitinating enzyme USP9X and the radiosensitivity of human glioma U251by measure the expression of USP9X and β-catenin under different does of radiotherapy.
Methods glioma cultured U251cells treated with0-10Gy radiation, measure USP9X and β-catenin on both gene and protein level by RT-PCR and Western-blot, and FCM is used to evaluate apoptosis.
RESULTS Under1Gy radiation, the express level of USP9X and β-catenin of U251in both gene and protein is higher than negetive control group. As the radio does increase from2Gy to10Gy, the expression of USP9X and β-catenin is lower down,together with radio dose. The apoptotic percentage of U251cell has a positive relationship with radio does.
Concusions Under1Gy Raidiation, the expression level of USP9X and β-catenin is higher than any other group, which proves low does of radiation can improve the radio-resistance of U251cell and increase of DNA repair. As radiation does increases, the express of USP9X and β-catenin is reduced, showes that high does of radiotherapy induce U251cell not able to complete DNA rapair and increase cell apoptosis. Above all,USP9X has a close connection with apoptosis of glioma cell after radiation.
引文
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[8]Giles RH, van Es JH, Clevers H. Caught up in a Wnt storm:Wnt signaling in cancer[J]. Neurosurg Focus,2006,20(4):El
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[14]Luo W, Peterson A, Garica BA, et al. Protein phosphatese I regulates assembly and function of the beta-catenin degradation complex. EMBO J,2007,26(6):1511-1521
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