目的 研究自噬与人鼻咽癌CNE-2细胞放射敏感性之间的关系.方法 采用慢病毒介导的RNA干扰技术建立稳定沉默自噬相关基因ATG5的人鼻咽癌CNE-2细胞系,实验分为未转染的CNE-2细胞组(对照组)、转染NC-shRNA的CNE-2细胞组(NC组)及转染ATG5-shRNA的CNE-2细胞组(ATG5组),应用CCK-8法、流式细胞术及克隆形成实验检测细胞增殖、凋亡及放射敏感性的变化.结果 CCK-8实验结果显示,与对照组和NC组相比,各剂量点ATG5组的细胞存活率均显著降低(F =3.755、46.086、8.609、44.160,P<0.05),绘制细胞生存曲线可见下调ATG5的表达后可以增加CNE-2细胞的放射敏感性;流式细胞术结果显示,经6 GyX射线照射后,ATG5组细胞的凋亡率较NC组及对照组明显升高(F =394.876,P<0.05);克隆形成实验结果提示沉默ATG5基因可增加鼻咽癌CNE-2细胞的放射敏感性.结论 降低鼻咽癌CNE-2细胞的自噬活性可以增强其放射敏感性.
Objective To investigate the effect of autophagy in radiosensitivity of nasopharyngeal carcinoma CNE-2 cells.Methods The expression of ATG5 in CNE-2 cells was inhibited by lentiviral mediated RNA interference.The cells were divided into three groups:control group,NC group and ATG5 group.Cell proliferation,apoptosis and radiosensitivity were detected by CCK-8 method,flow cytometry and colony-forming assay,respectively.Results Compared with the NC group and control group,the survival of ATG5-interfected cells was reduced (F =3.755,46.086,8.609,44.160,P 〈 0.05).After 6 Gy X-ray irradiation,the apoptosis index of the ATG5 group significantly higher than that of NC group and control group (F =394.876,P 〈 0.05).In addition,the radiosensitivity of ATG5 group was also higher than that of control cells.Conclusions Suppression of autophagy activity enhances the radiosensitivity of human nasopharyngeal carcinoma cells.