相同遗传背景不同辐射抗拒鼻咽癌细胞miRNA差异表达的研究

Relationship between radioresistance and microRNA in the same genetic background nasopharyngeal carcinoma cells

目的:在验证相同遗传背景鼻咽癌(NPC)细胞CNE-2R和CNE-2不同辐射抗拒性的基础上,探索微小RNA(miRNA)在CNE-2R和CNE-2中的表达差异与肿瘤辐射抗拒的关系。方法:通过观察X线照射对CNE-2R和CNE-2细胞克隆形成数目的影响,利用SigmaPlot软件进行分析及线性二次模型拟合存活曲线,比较CNE-2R和CNE-2细胞剂量存活曲线及其生物学参数;采用μParafloTM微流体芯片检测,用激光扫描仪(GenePix4000B,Molecular Device)采集杂交图像,经LOWESS滤器规范信号后分析数据差异,根据Targetscan311数据库资料(http://www.targetscan.org),预测CNE-2R和CNE-2细胞miRNA差异表达与NPC不同辐射抗拒的关系。结果:发现在CNE-2R和CNE-2细胞中存在miRNA的差异表达,即与CNE-2细胞比,在检测到的719个miRNA中,CNE-2R细胞中有37个上调,29个下调,其中检测量的绝对值≥2000且两者相差≥2倍的miRNA共12个:hsa-miR-200b、hsa-miR-224、hsa-miR-26b、hsa-miR-125a-5p、hsa-miR-205、hsa-let-7e、hsa-let-7g、hsa-miR-19b、hsa-miR-24、hsa-miR-103、hsa-miR-106b和hsa-miR-93。分析结果表明显著差异表达的miRNA与辐射抗拒密切相关。结论:相同遗传背景不同辐射抗拒NPC细胞株CNE-2R和CNE-2细胞的miRNA表达存在差异;差异表达的miRNA与辐射抗拒相关。

AIM： To discuss the relationship between the discrepancy of microRNA （miRNA） and radioresistance in nasopharyngeal carcinoma （NPC） cells CNE -2R and CNE -2 on the basis of validating their different radioresistance. METHODS： Following the exposure of X ray on the clones of CNE -2R and CNE -2 cells, the dose - survival curve and biological characteristics of CNE- 2R and CNE -2 cells were determined by SigmaPlot software and the linear quadratic model of survival curve analysis. MicroRNAs were detected by μParaflo^TM microfluidic chip, hybridization images collected by a laser scanner （GenePix 4000B, Molecular Device） and the signals normalized by a LOWESS filter. The re- lationship between the discrepancy of NPC radioresistance and the expression of miRNA was predicted according to Targetscan3.1 database （ http ：//www. targetscan, org） after analyzing the data. RFSULTS ： Compared to CNE - 2 cells, 37 miRNAs were gain -of- function and 29 miRNAs were loss -of- function in CNE -2R cells among 719 detected miR-NAs. 12 miRNAs that detective value was more than 2 000 and 2 folds than the other were hsa - miR - 200b, hsa - miR - 224, hsa - miR - 26b, hsa - miR - 125a - 5p, hsa - miR - 205, hsa - let - 7e, hsa - let - 7g, hsa - miR - 19b, hsa - miR- 24, hsa- miR- 103, hsa- miR- 106b and hsa- miR- 93. Data showed that the distinct discrepancy of miRNAs was related to radioresistance. CONCLUSION ： The discrepancy of miRNAs is present in different radioresistant NPC cell lines and related to radioresistance.