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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
引用本文:马嘉林,;郭亚,;曲颂,;赵伟,;李龄,;苏芳,;李烨,;葛莲英,;朱小东. 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[J]. 中国癌症防治杂志, 2014, 0(2): 121-126
作者姓名:马嘉林,  郭亚,  曲颂,  赵伟,  李龄,  苏芳,  李烨,  葛莲英,  朱小东
作者单位:[1]广西医科大学附属肿瘤医院放疗科,南宁530021; [2]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710049; [3]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西安交通大学第二附属医院肿瘤科,西安710049
基金项目: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)
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约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

Effect of silencing gene CDKN1A on radiosensitivity of radioresistant human nasopharyngeal carcinoma CNE-2R cell
Affiliation:MA Jia-lin, GUO Ya, QU Song, ZHAO Wei, LI Ling, SU Fang, LI Ye, GE Lian-ying, ZHU Xiao-dong (Department of Radiotherapy, Affiliated Tumor Hospital of Guangxi Medical University; Key Laboratory of High-Incidence-Tumor Prevention & Treatment Guangxi Medical University,Ministry of Education;1Graduate School of Guangxi Medical University,Nanning 530021,P.R.China; ZDepartment of Oncology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710049, P.R.China)
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.
Keywords:Nasopharyngeal neoplasm  Radioresistance  CDKN1A  Transfect
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