含CpG寡核苷酸在人神经母细胞瘤裸鼠移植瘤模型中的抗肿瘤效应  

Antitumor effect of CpG oligonucleotides on human neuroblastoma xenografts in nude mice

作　　者：姚强华 汤燕静 高丰厚[2] 汤静燕[1] 

机构地区：[1]上海交通大学医学院附属上海儿童医学中心血液/肿瘤科,上海200127 [2]上海交通大学医学院附属第三人民医院,上海201900

出　　处：《癌症》2009年第4期344-349,共6页Chinese Journal of Cancer

基　　金：上海市教委项目(No.05BZ48)~~

摘　　要：背景与目的:含有非甲基化CpG二核苷酸的寡脱氧核苷酸(CpG oligonucleotides,CpGODN)序列可以模拟细菌DNA特征,作为"异己信号"刺激机体免疫系统,发挥抗肿瘤作用。本研究探讨在人神经母细胞瘤(neuroblastoma,NB)裸鼠移植瘤模型中,CpGODN是否可以通过调节非特异性免疫来介导抗肿瘤效应。方法:体外WST-1实验明确CpGODN对神经母细胞瘤细胞株SK-N-MC的生长影响,建立NB荷瘤动物模型,当肿瘤生长到皮下可触及时,随机分组后给予不同注射处理(生理盐水、non-CpGODN、CpGODN)、观察各组小鼠肿瘤生长情况,病理组织学方法检测各组肿瘤组织的异同,免疫荧光组化法检测自然杀伤细胞(natural kill cell,NK)和巨噬细胞的浸润。结果:体外实验证明CpG ODN对SK-N-MC细胞无明显的直接细胞毒性及生长抑制作用。CpGODN处理组与non-CpG ODN、生理盐水组相比,肿瘤细胞生长明显受到抑制[(0.14±0.03)cm3vs.(2.97±0.40)cm3、(3.80±1.12)cm3(P<0.01)]。HE染色显示与肿瘤细胞生长活跃的对照组相比,CpG ODN处理组肿瘤组织中有较多的炎性细胞浸润及大片坏死灶。免疫荧光染色发现CpG ODN处理组与生理盐水组、non-CpG ODN组相比较瘤体内有明显的NK、巨噬细胞浸润。结论:具有免疫刺激作用的CpG ODN可能通过调节NK、巨噬细胞的活性来抑制NB生长,发挥抗肿瘤作用。Background and Objective. Synthetic CpG oligodeoxynucleotides （CpG ODN） containing unmethylated CpG dinucleotide motifs, which mimic the effects of bacterial DNA, can stimulate the host＇s immune defense to reject cancer cells as ＂non-self＂ signal. This study was to evaluate the antitumor effect of CpG ODN against human neuroblastoma xenografts in nude mice depending on the innate immunity. Methods. The cytotoxity of CpG ODN on neuroblastoma SK-N-MC cells was detected by WST-1 assay in vitro. Neuroblastoma xenografts were built subcutaneously in nude mice. When palpable tumor developed, the mice were randomized into normal saline （NS）, non-CpG ODN, and CpG ODN groups （each group contained six mice）, and were administered every other day for two weeks. When tumor grew to 5 cm^3, the mice were killed to observe tumor morphology by histology and histochemistry. Results： CpG ODN had no cytotoxicity on SK-N-MC cells in vitro, Tumor volume was significantly smaller in CpG ODN group than in NS and non-CpG ODN groups at the end of the observation [（0.14±0.03） cm^3 vs. （2.97±0.40） cm^3 and （3.80±1.12） cm^3, P〈0.01]. On HE-stained sections, the tumor tissues of CpG ODN group showed intratumoral infiltration of inflammatory cells and large areas of necrosis, whereas those of controls showed less infiltration and no necrosis. By immunohistochemistry, the tumor tissues of CpG ODN group showed more natural killer （NK） cells and macrophages as compared with those of control groups. Conclusion： CpG ODN may have therapeutic effect on neuroblastoma in nude mice via mediating the activity of NK cells and macrophages.

关 键 词：CpG ODN 脑肿瘤 神经母细胞瘤 移植瘤 小鼠 免疫刺激 NK细胞 巨噬细胞 非特异性免疫 

分 类 号：R739.44[医药卫生—肿瘤]