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目的:通过检测放疗对胶质瘤细胞β-catenin及去泛素化酶USP9X表达的影响,以期了解去泛素化酶USP9X与胶质瘤细胞放射敏感性的关系。方法:培养胶质瘤细胞U251细胞株,通过不同放疗剂量照射后,Realtime 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,流式细胞仪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的表达水平最高,随后随放疗剂量增加,USP9x和β-catenin基因表达成下降趋势,WesternBlotting结果显示USP9x、β-catenin的蛋白表达量与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放疗剂量照射时USP9X和β-catenin的表达水平最高,证明小剂量放疗可提高U251细胞的放射抵抗,随后随放疗剂量增加,USP9x和β-catenin的表达成下降趋势,证明USP9x与胶质瘤细胞的放射后凋亡水平相关。
OBJECTIVE: To investigate the effect of radiotherapy on the expression of β-catenin and the deubiquitinase USP9X in glioma cells in order to understand the relationship between the dextranase USP9X and the radiosensitivity of glioma cells. Methods: U251 glioma cell line was cultured. The expression of USP9X and β-catenin in cells was detected by Realtime PCR. The protein expression of USP9X and β-catenin was detected by Western Blotting. The expression of P-catenin was detected by flow cytometry The apoptosis of U251 cells treated with different dosages of radiation. Results: The results of Real Time PCR showed that the expression of USP9X and β-catenin were highest when irradiated with 1 Gy of radiotherapy, and then the expression of USP9x and β-catenin decreased with the increase of radiotherapy dose. Western Blotting showed that the protein of USP9x and β-catenin The expression level was positively correlated with the gene expression level. The apoptotic test results showed that the apoptotic rate was positively correlated with the radiation dose. CONCLUSIONS: The expression of USP9X and β-catenin is the highest at 1Gy irradiation dose, which shows that low dose radiation can increase the radiation resistance of U251 cells. With the increase of radiation dose, the expression of USP9x and β-catenin decrease. It is proved that USP9x and Glioma cells after radiation related to the level of apoptosis.