微小RNA-150对鼻咽癌细胞放疗抵抗的影响研究

发布时间：2018-01-25 12:42

本文关键词： 鼻咽肿瘤放射疗法微小RNA- 糖原合成酶激酶类　出处：《中国全科医学》2016年12期 　论文类型：期刊论文

【摘要】：目的探讨微小RNA(miR)-150对于鼻咽癌细胞放疗抵抗的影响,并分析其机制。方法 2014年1月—2015年6月,建立放疗抵抗的鼻咽癌细胞株CNE-2R。取CNE-2和CNE-2R,通过克隆形成实验计算不同放疗剂量下(0、3、6 Gy)的克隆形成率(PE)、细胞存活率,采用MTS法检测照射1、2、3、4、5 d时的细胞增殖活性,实时定量荧光聚合酶链式反应(Real-time PCR)法检测miR-150、糖原合成酶激酶3β(GSK3β)mRNA水平,Western blotting法检测GSK3β水平。将CNE-2随机分为miR-150模拟物组(转染miR-150模拟物)、微小RNAs(miRs)无关序列组(转染miRs无关序列),通过荧光素酶报告载体实验检测荧光素酶活性。结果 0 Gy放疗剂量时,CNE-2与CNE-2R的PE、细胞存活率比较,差异无统计学意义(P0.05);3 Gy、6 Gy放疗剂量时,CNE-2R的PE、细胞存活率高于CNE-2(P0.05)。照射1~5 d时,CNE-2R的细胞增殖活性高于CNE-2(P0.05)。CNE-2R中miR-150水平高于CNE-2,GSK3βmRNA水平低于CNE-2(P0.05)。CNE-2R中GSK3β水平低于CNE-2(P0.05)。转染GSK3β野生型质粒后,miR-150模拟物组荧光素酶活性低于miRs无关序列组(P0.05);转染GSK3β突变型质粒后,miRs无关序列组与miR-150模拟物组荧光素酶活性比较,差异无统计学意义(P0.05)。结论miR-150参与了鼻咽癌细胞的放疗抵抗,GSK3β是miR-150的直接靶基因,miR-150-GSK3β轴可能成为一种新的用于合理治疗鼻咽癌的策略。
[Abstract]:Objective to study the effect of RNA (miR) -150 for radiotherapy of nasopharyngeal carcinoma cells resistance, and analyze its mechanism. Methods from January 2014 to June 2015, the establishment of radioresistance of nasopharyngeal carcinoma cell line CNE-2R. CNE-2 and CNE-2R, through cloning experiment calculation different radiotherapy dose (0,3,6 Gy) the clone formation rate (PE), the cell survival rate. MTS method was used to detect 1,2,3,4,5 irradiation when D cell proliferation, real-time quantitative polymerase chain reaction (Real-time PCR) method to detect miR-150, glycogen synthase kinase 3 beta (GSK3 beta) mRNA level detection of GSK3 beta Western level blotting method. The CNE-2 were randomly divided into miR-150 group (transfected with miR-150 mimic analogue), tiny RNAs (miRs) group (transfected with miRs sequence independent, unrelated sequence) by luciferase reporter vector assay of luciferase activity. Results of the 0 Gy radiation dose, CNE-2 and CNE-2R PE, the cell survival rate than Relatively, the difference was not statistically significant (P0.05); 3 Gy, 6 Gy dose, CNE-2R PE, the cell survival rate was higher than that of CNE-2 (P0.05). 1~5 D irradiation, cell proliferation activity of CNE-2R was higher than that of CNE-2 (P0.05) miR-150 level in.CNE-2R is higher than CNE-2, GSK3 beta mRNA level was lower than that of CNE-2 (P0.05) GSK3.CNE-2R beta level is lower than CNE-2 (P0.05). The wild type plasmid after transfection of GSK3 beta, miR-150 analogue group luciferase activity was lower than that of miRs group (P0.05); sequence independent transfection of GSK3 beta mutant plasmid, miRs group and unrelated sequence analogue of miR-150 luciferase activity were compared, the difference was not statistically significant (P0.05) conclusion. MiR-150 is involved in the radiotherapy of nasopharyngeal carcinoma cells are resistant to GSK3 beta is a direct target gene of miR-150, miR-150-GSK3 beta axis may become a new strategy for rational treatment of nasopharyngeal carcinoma.

【作者单位】：南华大学附属第二医院耳鼻咽喉头颈外科;南华大学肿瘤研究所;
【基金】：国家自然科学基金资助项目(81172210) 湖南省省级科技计划项目(2014SK3081)
【分类号】：R739.63
【正文快照】： Influence of miR-150 on the Radioresistance of Nasopharyngeal Carcinoma Cells ZHANG Xian-feng,HUANG Yuan-jian,XIAO Juan,et al.Department of Otorhinolaryngology,the Second Hospital,University of South China,Hengyang421001,China鼻咽癌是一种具有较强局部侵袭

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