下调miR-210增加放射抗拒鼻咽癌细胞系CNE-2R的放射敏感性

Down-regulation of miR-210 enhances radiosensitivity in radio-resistant nasopharyngeal carcinoma cell line CNE-2R

目的研究下调miR-210对鼻咽癌放射抗拒细胞的放射敏感性的影响。方法用剂量梯度法建立鼻咽癌放射抗拒细胞CNE-2R。用microRNA基因芯片检测CNE-2和CNE-2R细胞的miRNAs,用q PCR验证miR-210的相对表达量。用LV-hsa-miR-210-inhibition慢病毒感染CNE-2R细胞,q PCR检测鼻咽癌细胞系210-inhibition中miR-210的相对表达量,用FCM测CNE-2R和210-inhibition细胞的凋亡和周期,用克隆形成实验测CNE-2R和210-inhibition的存活分数。结果诱导成功的CNE-2R与CNE-2细胞比较,有93个表达差异>2倍的miRNAs。下调CNE-2R中的miR-210后,与CNE-2R比较,210-inhibition凋亡比例明显增加(P<0.05),处于G2/M期的比例明显增加(P<0.05),S期的比例明显减少(P<0.05),210-inhibition细胞较CNE-2R细胞的存活分数降低。结论下调miR-210可增加鼻咽癌放射抗拒细胞的放射敏感性,其可能作为治疗放射抗拒的新靶点。

Objective To explore the effect of miR-210 down-regulation on radiosensitivity of radio-resist- ant nasopharyngeal carcinoma cells. Methods The radio-resistant nasopharyngeal carcinoma cells （ CNE- 2 R） were prepared by dose gradient method. The miRNA mieroarray was used for detecting miRNA expres- sion profiles of the CNE-2 cells and CNE-2R cells, and miR-210 was verified by qPCR. After LV-hsa- miR-210-inhibition infection, the miR-210 in 210-inhibition cells was verified by qPCR. Apoptosis and cell cycle of 210-inhibition and CNE-2R cells were examined by flow cytometry, the survival fraction of cells was detected by clone formation assay. Results There were 93 miRNAs molecules with expression re- markable change over 〉 2 fold in CNE-2R cell lines, which had a higher apoptosis rate than CNE-2R cells （P 〈 0.05 ）. The proportion of 210-inhibition cells in the G2/M phase was more than in the CNE-2R cells （ P 〈0.05） , the S phase of they were less than in the CNE-2R cells（P 〈0.05）. The survival fractions of 210-inhibition cells significantly reduced as compared with CNE-2R cells. Conclusions Down-regulation of microRNA-210 can enhance radiosensitivity in radio-resistant nasopharyngeal carcinoma cells, which might be a new target for the management of radiation resistance.