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中国癌症防治杂志

中国癌症防治杂志 ›› 2014, Vol. 6 ›› Issue (2): 121-126.doi: 10.3969/j.issn.1674-5671.2014.02.05

• 基础研究 • 上一篇    下一篇

沉默CDKN1A基因对鼻咽癌放射抗拒性CNE-2R细胞放射敏感性的影响

  

  1. 广西医科大学附属肿瘤医院放疗科；广西医科大学区域性高发肿瘤早期防治研究教育部重点实验室；广西医科大学研究生学院；西安交通大学第二附属医院肿瘤科
  • 出版日期:2014-06-25 发布日期:2014-07-08
  • 通讯作者: 朱小东 E-mail:zhuxiaodong83@163.com
  • 基金资助:

    国家自然科学基金资助项目（30860329）；广西自然科学基金资助项目（桂科自0832229）；广西高等学校重大科研基金资助项目（201101D004）；广西卫生厅科研基金资助项目（重2011076）

Effect of silencing gene CDKN1A on radiosensitivity of radioresistant human nasopharyngeal carcinoma CNE-2R cell

  • Online:2014-06-25 Published:2014-07-08

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被引次数

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摘要： 目的 探讨CDKN1A基因的表达与鼻咽癌放射敏感性的关系。方法 构建慢病毒表达载体LV-CDKN1A-RNAi并转染鼻咽癌放射抗拒性CNE-2R细胞，设转染LV-CDKN1A-RNAi慢病毒的CNE-2R细胞为实验组，转染阴性对照慢病毒的CNE-2R细胞为阴性对照组，未转染的CNE-2R细胞为空白对照组。用CCK-8法、细胞克隆形成实验及流式细胞术分别检测各组细胞增殖、放射敏感性及细胞周期的变化。结果 成功构建了CDKN1A基因沉默的CNE-2R细胞，CCK-8法检测显示实验组CNE-2R细胞在照射6 Gy后生长受到抑制，且随时间延长其抑制作用更为明显。细胞克隆形成实验显示实验组CNE-2R细胞放射敏感性增强（放射增敏比为SER=1.24）。流式细胞术检测显示实验组与对照组细胞相比，G0 /G1期和G2 /M期细胞分布在X射线照射6 Gy前后明显改变（P＜0.05）。结论 CDKN1A基因沉默能增强鼻咽癌放射抗拒性CNE-2R细胞的放射敏感性，CDKN1A基因的表达可能与鼻咽癌放射敏感性相关，有望成为鼻咽癌治疗的新靶点。

关键词: 鼻咽肿瘤, 放射敏感性, CDKN1A, 转染

Abstract: Objective To investigate the relationship between the CDKN1A gene expression and the change of radiosensitivity of human nasopharyngeal carcinoma. Methods Lentiviralvector-mediated RNA interference（LV-CDKN1A-RNAi） targeting was constructed and transfected it to CNE-2R cells.CNE-2R cells transfected with LV-CDKN1A-RNAi were taken as experimental group，those transfectedwith control virus and without any treatment were taken as control group and blank group.The proliferation inhibitory rate was measured by CCK-8 assay.Colony formation assay was performed to determine the radiosensitizing effect.Cell cycle distribution was analyzed by flow cytometry. Results The stably silenced expression of CDKN1A CNE-2R cell line was established.CCK-8 assay showed that the growth of CNE-2R cell of the experimental group was inhibited after 6 Gy irradiating，which was closely related to growth time.The results of colony formation assay showed that the radiosensitivity of CNE-2R cell of the experimental group was more enhance（SER=1.24） than the other groups.Comparing with control group，cell cycle distribution showed that the cell distribution of G0 /G1 and G2 /M cycle in the experiment group was significantly changed before and after 6 Gy irradiating. Conclusions Silencing gene CDKN1A may enhance the radiosensitivity of CNE-2R cells of human nasopharyngeal carcinoma，which maybe used as a thera-peutic target for nasopharyngeal carcinoma.

Key words: Nasopharyngeal neoplasm, Radioresistance, CDKN1A；Transfect