HMGB1在多形性成胶质细胞瘤中的作用及其机制研究  

The role of HMGB1 in glioblastoma mutiforme and its mechanism study

作　　者：李文涛[1] 魏艳[1] 任春莹 王建峰[1] 洖咏 罗文红[1] 姚妮[1] 王颖[1] 孟喜军[1] 

机构地区：[1]西安交大第一附属医院神经外科,陕西西安710061

出　　处：《临床和实验医学杂志》2017年第16期1596-1600,共5页Journal of Clinical and Experimental Medicine

摘　　要：目的研究高迁移率蛋白(HMGB1)在多形性成胶质细胞瘤(GBM)中的作用及其机制。方法用9 L胶质瘤细胞颅内接种Wistar鼠构建同源GBM模型。建立GBM模型鼠后将其随机分为3组:GBM+Saline(n=20),GBM+Ad(n=20),GBM+Ad+Gly(n=20)。10 d后对GBM+Ad组鼠进行Ad-TRAIL感染(8×107pfu/5μl),GBM+Ad+Gly组鼠进行Ad-TRAIL感染和甘草酸(Glycyrrhizin)喂养(稀释于氢氧化钠,100 mg腹腔注射,每天两次连续喂养10d),GBM+Saline组和对照组用生理盐水处理。采用ELISA测定体内和体外HMGB1的含量水平,TUNEL法测定细胞的凋亡,免疫细胞组化(ICC)检测HMGB1的表达。结果 HMGB1在GBM+Ad组中的血清含量为5.540±0.054 ng,显著高于GBM+saline组的0.0060±0.004 ng(P=0.0001);GBM+Ad+Gly组中HMGB1血清含量(0.07±0.016 ng)显著低于GBM+Ad组(P=0.04);GBM+saline组的HMGB1血清含量高于对照组,但差异无统计学意义(P=0.923)。免疫细胞组化实验中,HMGB1在GBM-Ad组中呈高阳性表达。体外检测中,GBM+Ad组中HMGB1浓度(77.505±1.196 ng)高于GBM+saline组(8.623±0.052 ng)(P=0.0003);GBM+Ad0组中HMGB1浓度低于GBM+saline组,但差异无统计学意义(P>0.05)。TUNEL法检测Ad-TRAIL感染GBM原代细胞后的细胞凋亡发现,GBM+Ad组的细胞死亡率为(78.91±0.17)%,显著高于GBM+saline组(18.29±0.73)%(P=0.04)。相关性分析结果显示HMGB1含量水平与细胞死亡率呈正相关,R2=0.9976,P<0.0001。GBM+saline和GBM+Ad+Gly组鼠在第12天开始出现死亡,在第24天和第25天全部死亡;而GBM+Ad组在第16天开始出现死亡,在第40天时有50%(5/10)长期存活。结论HMGB1在GBM治疗中高释放,且其浓度与细胞凋亡正相关。HMGB1是GBM的治疗靶点,并且是检测腺病毒治疗或免疫治疗GBM疗效的潜在生物标记。Objective To study the role of HMGB1 in glioblastoma multiforme（ GBM） and its mechanism. Methods GBM rats model were made by intracerebral inoculation with 9 L glioma cells and then treated with Ad-TRAIL. The rats were randomly divided into three groups：GBM ＋ Saline（ n = 20）,GBM ＋ Ad（ n = 20）,GBM ＋ Ad ＋ Gly（ n = 20）. Ad-TRAIL infection（ 8 × 107pfu/5 μl） was performed on GBM ＋Ad mice,Ad-TRAIL infection and Glycyrrhizin feeding were performed in GBM ＋ Ad ＋ Gly group. The GBM ＋ Saline group and the control group were treated with physiological saline. s ELISA was used to test HMGB1 concentration in vitro and in vivo. TUNEL assay was performed for cell apoptosis test and immunocytochemistry was conducted for HMGB1 detection. Results Serum level of HMGB1 in GBM ＋ Ad group was 5. 540± 0. 054 ng,which was significant higher than that in GBM ＋ saline group（ 0. 0060 ± 0. 004 ng）,P = 0. 0001; Serum level of HMGB1 in GBM＋ Ad ＋ Gly group（ 0. 07 ± 0. 016 ng） was less than that in GBM ＋ Ad group and P = 0. 04. Serum level of HMGB1 in GBM ＋ saline group was higher than that in control group,while P = 0. 923 and ther was no significant difference. GBM-Ad rats showed high HMGB1 positive measured by immunocytochemistry. The concentration of HMGB1 in GBM ＋ Ad cell supernatant（ 77. 505 ± 1. 196 ng） was obviously higher than that in GBM ＋ saline（ 8. 623 ± 0. 052 ng）（ P = 0. 0003）. GBM ＋ Ad0 group was less than GBM ＋ saline,but with no difference（ P 〉 0. 05）. In cell apoptosis test of primary GBM cell treated with Ad-TRAIL,cell death from GBM ＋ Ad group was（ 78. 91 ± 0. 17） %,which was significantly higher than that from GBM ＋ saline group（ 18. 29 ± 0. 73） %（ P = 0. 04）. HMGB1 had a positive correlation with cell death（ R2= 0. 9976,P〈 0. 0001）. At day 12,rat death appeared in GBM ＋ saline and GBM ＋ Ad ＋ Gly groups and all rats were died in these two groups at day 24 and25,respectively; however,rat death happened at

关 键 词：WISTAR鼠 多形性成胶质细胞瘤 高迁移率蛋白 肿瘤坏死因子相关性凋亡诱导配体 甘草酸 

分 类 号：R739.41[医药卫生—肿瘤]