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5-氮杂胞苷增加鼻咽癌细胞株C666-1放射敏感度的实验

5-azacytidine Increases Radiation Sensitivity of Nasopharyngeal Carcinoma Cell Line C666-1

  • 摘要:
    目的  研究5-氮杂胞苷(5-azacytidine, 5-Aza)对人鼻咽癌细胞C666-1放射敏感度的影响及可能机制。
    方法 用不同浓度5-氮杂胞苷(0、500、1000、3000、5000 nmol/L)处理C666-1细胞,MTT法检测C666-1细胞增殖,流式细胞仪检测C666-1细胞周期,Western blot法检测细胞凋亡相关蛋白cleaved Caspase-3、cleaved PARP及DNA损伤相关蛋白γ-H2AX的表达。
    结果 5-氮杂胞苷随着药物浓度的增加,对C666-1细胞生长抑制作用增强。用1 000 nmol/L 5-氮杂胞苷处理鼻咽癌细胞进行放射增敏研究。鼻咽癌细胞C666-1接受单纯电离辐射时SF2(照射2 Gy时的细胞存活率)为48%,5-氮杂胞苷联合电离辐射时SF2降为31%,放射增敏比为1.37。5-氮杂胞苷能增加电离辐射诱导凋亡相关蛋白cleaved Caspase-3及cleaved PARP的表达。
    结论 5-氮杂胞苷增加鼻咽癌C666-1细胞对电离辐射的敏感度,可能的机制为5-氮杂胞苷能增加电离辐射诱导细胞凋亡。

     

    Abstract:
    Objective To investigate the effect of 5-azacytidine (5-Aza) on the radiation sensitivity of human nasopharyngeal carcinoma cell line C666-1 and its underlying mechanism.
    Methods C666-1 cells were treated by 5-azacytidine with different concentrations (0, 500, 1000, 3000, 5000nmol/L). Cell growth was determined by MTT, cell cycle was detected using flow cytometry, and the expression of apoptosis-related proteins, such as cleaved Caspase-3, cleaved PARP and DNA damage related protein gamma H2AX were assayed by Western blot.
    Results With the increase of drug concentration, 5-azacytidine enhanced the inhibition of C666-1 cells growth gradually. We further studied the radiation sensitivity of nasopharyngeal carcinoma cells using 1000nmol/L 5-azacytidine. SF2 (cell survival rate after radiation of 2Gy) of C666-1 cells after receiving radiation alone was 48%, SF2 was decreased to 31% after treated by 5-azacytidine combined radiation, and the radiotherapy sensitization ratio was 1.37. 5-azacytidine increased the expression of apoptosis-related proteins cleaved Caspase-3 and cleaved PARP.
    Conclusion 5-azacytidine could enhance the radiation sensitivity of nasopharyngeal carcinoma C666-1 cells and the possible mechanism is that 5-azacytidine may increase the radiation-induced apoptosis.

     

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