3MA抑制自噬降低头颈鳞癌细胞放疗抵抗性的实验研究  

Study on 3MA reduce radioresistance in squamous cell carcinoma of the head and neck via autophagy inhibition

作　　者：敬前程[1] 刘勇[2] 李果[2] 卢善翃 刘超[2] 黄东海[2] 周芳[1] 邱元正[2] JING Qiancheng;LIU Yong;LI Guo;LU Shanhong;LIU Chao;HUANG Donghai;ZHOU Fang;QIU Yuanzheng(Department of Otolaryngology Head and Neck Surgery,Changsha Central Hospital,University of South China,Changsha,Hunan,410004,China;Department of Otolaryngology Head and Neck Surgery,Key Laboratory of Otolaryngology Major Disease Research of Hunan Province,Xiangya Hospital,Central South University,Changsha,Hunan,410008,China)

机构地区：[1]南华大学附属长沙市中心医院耳鼻咽喉头颈外科,湖南长沙410004 [2]中南大学湘雅医院耳鼻咽喉头颈外科耳鼻咽喉重大疾病研究湖南省重点实验室,湖南长沙410008

出　　处：《中国耳鼻咽喉颅底外科杂志》2020年第5期532-536,共5页Chinese Journal of Otorhinolaryngology-skull Base Surgery

基　　金：国家自然科学基金(81602684,81773243)。

摘　　要：目的探讨6-氨基-3-甲基嘌呤(3MA)对头颈鳞癌细胞放疗抵抗性的影响。方法蛋白印迹法(Western blotting)检测CNE2、6-10B及其放疗抵抗细胞CNE2-Rs、6-10BRs中LC3B蛋白的表达;3MA作用于CNE2-Rs及TU686细胞后LC3B蛋白的表达;克隆集落形成实验检测3MA对头颈鳞癌细胞的放疗抵抗能力及生存分数的改变;细胞免疫荧光法检测3MA对放疗致DNA双链损伤相关指标γ-H2AX焦点数量的改变。结果 LC3B蛋白在放疗抵抗细胞较亲本细胞中表达增加;3MA能有效抑制自噬膜蛋白LC3B的表达;3MA抑制自噬后CNE2-Rs及TU686细胞克隆集落形成能力减弱,生存分数降低;3MA抑制自噬后CNE2-Rs及TU686细胞放疗组中γ-H2AX焦点数量较对照组显著增加。结论 3MA抑制自噬后可能影响放疗后DNA双链损伤修复,进而降低头颈鳞癌细胞的放疗抵抗性。Objective To study the function of 3 MA on the radioresistance in squamous cell carcinoma of the head and neck.Methods Western blotting was used to detect the LC3 B protein in CNE2,6-10 B and its radioresistance cell lines CNE2-Rs,6-10 BRs, in CNE2-Rs and TU686 cells after 3 MA treatment. Colony formation assay was applied to check the alteration of radioresistant capacity and survival fraction.Immunofluorescence was done to detect the expression of γH2 AX foci,which was a surrogate marker of DNA doublestrand breaks.Results The expression of LC3 B protein was increased in radioresistant cell lines and inhibited in 3 MA treatment groups.3 MA weakened the colony formation ability and decreased the survival fraction.Also,3 MA significantly increased the γH2 AX foci formation compared with the control group after irradiation.Conclusion The inhibition of autophagy by 3 MA may affects the repair of DNA double strand damage after radiotherapy, thereby reducing the radioresistance of squamous cell carcinoma of head and neck.

关 键 词：自噬 头颈鳞癌 放疗抵抗 3MA Γ-H2AX 

分 类 号：R739.91[医药卫生—肿瘤]