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• W3013402402 abstract "Objective:To investigate the effect of Notch1 gene on radiosensitivity of nasopharyngeal carcinoma cells and its molecular mechanism. Method:A Notch1-knockout CNE-2 cell line was constructed using CRISPR/Cas9 system, and the expression of Notch1 gene was detected by RT-PCR and Western blot. After treatment with different doses of radiation, the survival fraction （SF） of each group was calculated, and used the GraphPad Prism 6.0 software and the Linear quadratic model were used to calculate the fitted dose survival curve and the sensitivity enhancement ratio（SER）. Taking 6 Gy as radiation dose, the experiment was divided into four groups: Notch1（+） group, Notch1（-） group, IR+Notch1（+） group and IR+Notch1（-） group. CCK-8 assay was used to detect cell proliferation in each group. Annexin V-FITC/PI double staining assay was used to detect the changes of apoptosis in each group. The expression of H2AX, CyclinD1, Bax, Bcl-2 and GAPDH proteins were detected by Western blot. Result:The CNE-2 cell line with Notch1 gene knockout was successfully constructed. The clonogenic assay showed knockout of Notch1 enhanced the radiosensitivity of NPC cells. The CCK-8 assay showed that cell proliferation and cell viability were significantly reduced in the IR+Notch1（-） group compared with the IR+Notch1（+） group（P<0.05）. Annexin V-FITC/PI double staining assay showed that the IR+Notch1（-） group had the highest apoptosis rate compared with the other groups （P<0.05）. Western blotting demonstrated that the expression of γH2AX was significantly increased after irradiation of Notch1 nasopharyngeal carcinoma cells, the expression of Cyclin-D1 was increased, and the ratio of Bax:Bcl-2 was higher. Conclusion:Knockout of Notch1 signaling molecule can effectively improve the radiosensitivity of NPC cells cultured in vitro, which may be a potential target for radiosensitization of NPC.目的：探讨Notch1基因对鼻咽癌细胞放疗敏感性的影响及其分子机制。 方法：使用CRISPR/Cas9技术构建敲除Notch1基因的CNE-2细胞（人鼻咽癌细胞株），通过RT-PCR以及Western blot法检测Notch1基因的表达。经不同剂量射线照射处理后，通过计算各组存活分数（SF），使用Linear-quadratic模型计算拟合剂量生存曲线，计算放射增敏比（SER）。以6 Gy为照射剂量，将实验分为4个组：Notch1（+）组、Notch1（-）组、IR+Notch1（+）组和IR+Notch1（-）组。CCK-8法检测各组细胞增殖情况，Annexin V-FITC/PI双染法检测各组细胞凋亡变化，Western blot检测各组细胞γH2AX、CyclinD1、Bax、Bcl-2和GAPDH蛋白的表达差异。 结果：敲除Notch1基因的CNE-2细胞系构建成功。CCK-8实验显示，敲除Notch1可提高鼻咽癌放疗敏感性：与IR+Notch1（+）组细胞相比，IR+Notch1（-）组细胞增殖和细胞存活率显著降低（P<0.05）。Annexin V-FITC/PI双染法显示，与其他组比较，IR+Notch1（-）组凋亡率最高（P<0.05）。Western blot结果显示，敲除Notch1的鼻咽癌细胞经过照射后，γH2AX的表达量明显升高，Cyclin D1的表达量增高，Bax：Bcl-2比值较高。 结论：敲除Notch1信号分子可有效提高体外培养的鼻咽癌细胞的放疗敏感性，其可能是鼻咽癌放疗增敏的一个潜在靶点。." @default.
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• W3013402402 date "2020-01-01" @default.
• W3013402402 modified "2023-09-26" @default.
• W3013402402 title "[Knockout notch1 gene can enhanced radiosensitivity of nasopharyngeal carcinoma cells]." @default.
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• W3013402402 doi "https://doi.org/10.13201/j.issn.1001-1781.2020.01.016" @default.
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