中文摘要：

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（P〉0.05）;3 Gy、6 Gy放疗剂量时,CNE-2R的PE、细胞存活率高于CNE-2（P〈0.05）。照射1-5 d时,CNE-2R的细胞增殖活性高于CNE-2（P〈0.05）。CNE-2R中miR-150水平高于CNE-2,GSK3βmRNA水平低于CNE-2（P〈0.05）。CNE-2R中GSK3β水平低于CNE-2（P〈0.05）。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（P〈0.05）;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（P〉0.05）。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。

英文摘要：

Objective To investigate the effect of miR-150 on the radioresistance of nasopharyngeal carcinoma （NPC）cells and analyze its mechanism. Methods From January 2014 to June 2015,we established a radioresistant NPC cell line CNE - 2R. CNE - 2 and CNE - 2R were taken,and clone formation assay was conducted to calculate cloning formation efficiency（PE）and cell survival rate with different radiotherapy doses（0,3 and 6 Gy）. MTS method was employed to detect cell proliferation activity on day 1,day 2,day 3,day 4 and day 5. Real - time PCR method was used to detect the levels of miR-150 and GSK3β mRNA,and GSK3β level was measured by Western blotting method. CNE - 2 cells were randomly divided into miR-150 mimics group（ transfection of miR-150 mimics） and miRs irrelevant sequence group（ transfection of miRs irrelevant sequence）,and luciferase activity was detected by luciferasereport carrier experiment. Results CNE - 2 and CNE -2R were not significantly different in PE and cell survival rate when the radiation dose was 0 Gy（P 〉 0. 05）. CNE - 2R was higher than CNE - 2 in PE and cell survival rate when the radiation doses were 3 Gy and 6 Gy（P 〈 0. 05）. After radiation for 1 day to 5 days,the activity of CNE - 2R cell proliferation activity was higher than CNE - 2（P 〈 0. 05）. CNE - 2R was higher in miR-150 and lower in GSK3β mRNA than CNE - 2（P 〈 0. 05）. CNE - 2R was lower than CNE - 2 in the level of GSK3β（P〈 0. 05）. After the transfection of GSK3β wild type plasmid,miR-150 mimics group was lower than miRs irrelevant sequence group in luciferase activity（P 〈 0. 05）;after the transfection of GSK3β mutant plasmid,miRs irrelevant sequence group and miR-150 mimics group were not significantly different in luciferase activity（P 〉 0. 05）. Conclusion miR-150 is involved in the radioresistance of NPC cells. GSK3β is a direct target gene of miR-150,and miR-150 - GSK3β axis may be a possible strategy for rational NPC treatment.