自噬在塞来昔布对脑胶质瘤SHG-44细胞放射增敏效应中的作用  

Effects of autophagic cell death in the role of celecoxib' radiosensitizing on human glioma SHG-44 cell line

作　　者：聂斌[1] 周菊英[2] 吴琼[2] 王利利[2] 徐晓婷[2] 

机构地区：[1]南京医科大学附属常州第二人民医院放疗科,江苏常州213003 [2]苏州大学附属第一医院放疗科,江苏苏州215006

出　　处：《苏州大学学报（医学版）》2012年第1期18-22,62,共6页Suzhou University Journal of Medical Science

基　　金：2006年江苏高校省级重点实验室开放研究课题

摘　　要：目的探讨自噬在塞来昔布对脑胶质瘤SHG-44细胞放射增敏中的作用。方法选择脑胶质瘤细胞株SHG-44作为实验对象,并分成对照组(C组)、塞来昔布组(D组)、辐射组(R组)和联合组(D+R组)。集落形成法检测塞来昔布对SHG-44细胞的放射增敏作用。吖啶橙染色、FITC标记LC3-Ⅱ抗体和电镜共同检测细胞的自噬水平。流式细胞仪检测细胞周期分布和凋亡情况。结果塞来昔布(30μmol/L)增加SHG-44细胞的放射敏感性,诱导肿瘤细胞自噬和G2/M期阻滞。电镜观察发现塞来昔布、辐射及联合作用后胞质内可见自噬体,而无明显的核浓缩和核碎片。吖啶橙染色和FITC-MAP1-LC3-Ⅱ标记自噬体发现D+R组细胞自噬水平明显高于R组(P<0.05)。D+R组G2/M期细胞比例为(34.26±2.20)%,R组为(29.15±1.99)%,D+R组明显多于R组(P<0.05),而相应的凋亡比例分别为(9.7±1.24)%和(8.2±0.93)%,两组差异无统计学意义(P>0.05)。随着辐射剂量的增加,细胞自噬水平增高,而细胞存活率呈指数性递减(决定指数R=0.98,P<0.05)。结论塞来昔布增强辐射诱导SHG-44细胞G2/M期阻滞、促进细胞自噬、诱导细胞自噬性死亡,可能是其增加放射敏感性的机制之一。Objective To analyze the effect of autophagy on celecoxib radiosensitizing human glioma SHG-44 cell line.Methods SHG-44 cells were divided into four groups：（1）control group（C）;（2）celecoxib only（D）;（3） radiation only（R）;（4）celecoxib plus radiation（D＋R）.Celecoxib＇s radiosensitization was measured by clongenic assay.The cell cycle redistribution and apoptosis were analyzed by flow cytometric analysis.Meanwhile,acridine orange（AO）,FITC-LC3-II antibody and TEM were used to detect autophagy.Results Celecoxib（30 μmol/L） radiosensitized SHG-44 cell by induced autophagy and cells G2/M arrest.Two days after 8 Gy irradiation or（and） celecoxib 30 μmol/L,SHG-44 cells showed by TEM prominent formation of autophagic vesicles with lamellar structures or residual digested material without nucleus condensation or segmentation.Next,cells were incubated by acridine orange or LC3-II antibody after irradiation or（and） celecoxib.The level of autophagy of（D＋R） group was higher than that in the R alone（P0.05）.The proportion of G2/M was（34.26±2.20）% of（D＋R） treatment,compared with 8 Gy irradiation alone（29.15±1.99）%（P0.05）.However,apoptotic cell death was（9.7±1.24）% and（8.2±0.93）% respectively without apparent statistic significance（P0.05）.As the level of autophagy increased,the clongenic survival decreased exponentially（correlation index R=0.98,P0.05）.Conclusion Celecoxib enhances SHG-44 cells G2/M phase arrest by radiation-induced and promoted cell autophagy.To induce autophagic cell death may be one of the mechanisms of celecoxib radiosensiting glioma cells SHG-44.

关 键 词：自噬 脑胶质瘤 放射疗法 塞来昔布 凋亡 

分 类 号：R730.264[医药卫生—肿瘤]