干扰CDC25A基因对CNE2细胞连续照射期间增殖的影响  

Effect of CDC25A gene knocked-down on the accelerated proliferation of continunous irradiated nasopharyngeal carcinoma cell line CNE2

作　　者：曲颂[1] 朱小东[1] 李相德[1] 李龄[1] 黎丹戎[1] 张玮[1] 

机构地区：[1]广西医科大学附属肿瘤医院,南宁530021

出　　处：《山东医药》2011年第1期15-17,共3页Shandong Medical Journal

基　　金：广西医疗卫生重点科研课题(200866);广西2007年研究生科研创新资助项目(10598200702M019)

摘　　要：目的采用RNA干扰技术沉默CDC25A基因,观察其对鼻咽低分化鳞癌细胞株(CNE-2)在连续照射中后期加速增殖现象的影响。方法构建、培养出CDC25A基因沉默效果稳定的CNE2-psilence4.1-CDC25A,接受射线2Gy/d连续照射5 d,分别采用MTT法、流式细胞术检测细胞的生长增殖活性,并在mRNA及蛋白水平上应用RT-PCR、Western Blot法进行验证。结果 MTT法检测结果表明,CNE2-psilence4.1-CDC25A细胞存活分数(SF)在第5天达到高峰,CNE2-psilence4.1-Control细胞SF在第3天达到高峰。流式细胞术检测结果显示,CNE2-psilence-4.1-CDC25A细胞在照射第5天S期细胞比(SPF)及增殖指数(PI)为最高值,CNE2-psilence4.1-Control细胞在照射第3天SPF值最高,在照射第4天PI值最高;RT-PCR、Western Blot法进一步验证CNE2-psilence4.1-CDC25A细胞在照射第1、3、5天的CDC25A的表达与MTT法、流式细胞术检测结果基本吻合。结论与CNE2-psilencer4.1-Control细胞相比,基因沉默的CNE2-psilencer4.1-CDC25A细胞在连续分割照射期间,细胞的生长速度减慢、生长高峰延迟。Objective To investigate the status of proliferation in the CNE2 cell lines during irradiation with CDC25A gene knocked-down. Mechods CNE2-psilence4. 1-CDC25A with CDC25A gene knocked-down was constructed and cul- tured ; MTT and flow cytometry assay were performed to determine the dynamics proliferation of CNE2-psilence4.1 -CDC25A cell lines and control cen lines during 60Co-γ 2 Gy 5-daycontinuous fractionated irradiation, and RT-PCR and western blotting were performed to verify the interfering effectiveness of CDC25A gene in the constructed cell line before and after irradiation. Results Dynamic changes of CNE2-psilence4.1-CDC25A cell lines and control cell lines proliferation were detected by MTT and FCM assay during 60Co-γ, 2 Gy 5-day-continuous fractionated irradiation. MTT method showed that the survival fraction（SF） was the highest on the fifth day in the CNE2-psilenced4.1-CDC25A cell, and on the third day in the CNE2-psilence4.1-Control cell. FCM showed that the peak of CNE2-psilence4.1 cell lines proliferation occurred in the 5th day while the control cell lines occurred in the 3rd and 4th day. Conclusions Compared with the control cell lines, there is decreased proliferation and delayed growth peak occurred in CNE2-psilencer4.1-CDC25A cell lines.

关 键 词：RNA干扰 CDC25A CNE-2 加速再增殖 

分 类 号：R739.63[医药卫生—肿瘤]