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
聂斌 1周菊英 2吴琼 2王利利 2徐晓婷2
作者信息
- 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
- 折叠
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( 30 μmol/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.
Abstract
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 radiosen-sitization 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 (xmol/L) radiosensitized SHG-44 cell by induced autophagy and cells G2/M arrest. Two days after 8 Gy irradiation or (and) celecoxib 30 jimol/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(P<0.05). The proportion of G2/M was (34.26 ±2.20)% of (D +R) treatment, compared with 8 Gy irradiation alone (29. 15 ± 1. 99)% (P <0. 05). However, apoptotic cell death was (9.7 ±1.24)% and (8.2 ±0.93)% respectively without apparent statistic significance (P>0. 05 ). As the level of autophagy increased, the clongenic survival decreased exponentially ( correlation index R =0.98 ,P <0.05). 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Key words
autophagy/malignant glioma/radiation/celecoxib/apoptosis引用本文复制引用
出版年
2012
NETL
NSTL
维普
万方数据