C6胶质瘤荷瘤大鼠刀照射后胶质瘤细胞死亡方式实验研究  

Gamma knife treatment of rat's gliomas:Relationship of the cell apoptosis and necrosis with irritation dose

作　　者：王鹏[1] 于金明[2] 潘绵顺 方兴根 

机构地区：[1]天津医科大学肿瘤医院 [2]山东省肿瘤防治院放疗科 [3]安徽省合肥市脑科医院

出　　处：《中国实验诊断学》2008年第3期321-324,共4页Chinese Journal of Laboratory Diagnosis

基　　金：卫生部科研基金课题(课题编号WKJ2005-3-006)

摘　　要：目的采用放射线照射大鼠C6脑胶质瘤模型,分析放射线诱导胶质瘤细胞死亡方式的发生规律。方法选用24只生后5-6周雌性SD大鼠,购自安徽医科大学实验动物中心。大鼠C6胶质瘤细胞购自中科院细胞库。所有大鼠均采用立体定向法制作脑胶质瘤模型,即入脑皮质下5 mm注入4μl体外培养和传代后的C6细胞悬液。随机抽签法将大鼠分为9 Gy照射组1、2 Gy照射组和模型组,每组8只。前2组分别于造模后21 d行照射剂量分别为9Gy、12Gy伽玛刀治疗,模型组未给予照射。治疗后1周取大鼠肿瘤与脑组织交界处组织,采用流式细胞仪检测肿瘤细胞坏死率及凋亡率,HE染色观察组织学变化。结果纳入大鼠24只,模型组和实验组2分别有1只大鼠在麻醉意外中死亡,其余22只大鼠均进入结果分析。①肿瘤细胞坏死率及凋亡率:两剂量照射组肿瘤细胞凋亡率和坏死率均高于模型组,差异有统计学意义(P<0.05);9 Gy照射组肿瘤细胞的凋亡率高于12 Gy照射组,坏死率低于12 Gy照射组,差异有统计学意义(P<0.01)。②组织学变化:各组肿瘤与脑组织均可见肿瘤生长,伽玛刀两剂量组治疗后出现了肿瘤中心坏死,坏死区细胞稀疏,向外细胞渐增多,其间含有很多的破碎细胞,再向外则为密集的瘤细胞,并有细胞内、外的水肿和出血,同时也有不同程度的核固缩细胞,12 Gy照射组较9 Gy照射组坏死增加。结论伽玛刀可诱导脑胶质瘤模型大鼠肿瘤细胞凋亡和坏死,在治疗剂量范围内,随着剂量的增加肿瘤坏死有增加趋势。Objective To observe the mechisms of apoptosis and necrosis induced by gamma knife treatment of rat glioma models, analysis the relationship of the cell apoptosis and necrosis with irritation dose. Methods Twenty-four female specific pathogen ffee（SPF） Spraque-Dawley rats, weigting 180 - 210 g, aged 5 - 6weeks, were purchased from Experimental Animal Center, Anhui Medical College.Rat C6 glioma ceils were purchased from cell bank of CAS.Annexin V-FTTC Reagent Kit （Bendermedsystem.com） and flow cytometry （Becton Dichinson FACSCalibur） were provided.The experiment was conducted in the Cancer Hospital of Tianjin Medical University and Gamma knife center of Hefei Brain Hospital from December 2006 to May 2007. All the rats were inoculated with C6 glioma ceils, ie. Four pL of C6 glioma ceils suspension were inoculated into 5 mm under cortex of the rots. All the rots were randomized divided into model group , 9 Gy trcatment group and 12 Gy trcatment group. Six rats in each group. Cell apoptosis and necrosis of tissue between normal brain tissue and gliomas were observed by Flow Cytometry examination one week after irradiation, and pathological changes in tumor tissues were identified with HE staining. Results Twenty four rats were selected in the experiment, and 2 rats of the model group and 12 Gy treatment group died from anaesthesia accident .311e remaining 22 rats were entered the result analysis.① Cell apoptosis and necrosis：Apoptosis and necrosis were increased significantly more in both treatment groups than model group after gamma knife irritation, and there was significant difference （P 〈 0.05） ; Apoptosis was significantly more in 9 Gy treatment group than 12 Gy treatment group,and there was significant difference （P 〈 0.01）;Necrosis was significantly less in 9 Gy treatment group than 12 Gy treatment group, and there was significant difference （ P 〈 0.01 ）. ② Pathological changes： A marked decrease in cellularity and patches of necrosis with disperse extracellular hemorrhage

关 键 词：伽玛刀 脑胶质瘤 凋亡 坏死 

分 类 号：R739.4[医药卫生—肿瘤]