抑制MDSC对神经母细胞瘤荷瘤小鼠CXCL12/CXCR4表达影响及在免疫治疗中的作用  

Inhibition of effect of myeloid-derived suppressor cell upon the expressions of CXCL12 and CXCR4 and immunotherapy in neuroblastoma-loaded BALB/c mice

作　　者：徐伟立[1] 李索林[1] 李萌[1] 周辉[1] 耿娜[1] 杨晓锋[1] Xu Weili;Li Suolin;Li Meng;Zhou Hui;Geng Na;Yang Xiaofeng(Department of Pediatric Surgery,Second Hospital,Hebei Medical University,Shijiazhuang 050000, China)

机构地区：[1]河北医科大学第二医院小儿外科,石家庄050000

出　　处：《中华小儿外科杂志》2019年第1期58-63,共6页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金面上项目(81472503);河北省人才工程培养经费资助科研项目(A201500143).

摘　　要：目的了解抑制神经母细胞瘤(neuroblastoma,NB)荷瘤小鼠体内髓系抑制性细胞(myeloid-derived suppressor cell,MDSC)对趋化因子12(cxc chemokine receptor ligand 12,CXCL12)及趋化因子受体4(cxc chemokine receptor type 4,CXCR4)表达影响及其在免疫治疗中的作用,探索控制NB生长和转移的新方法。方法采用SK-N-SH细胞接种法建立神经母细胞瘤BALB/c荷瘤小鼠模型,成瘤后切取标本,HE染色观察肿瘤组织结构。将荷瘤小鼠按随机数字表法分为对照组、生理盐水(NS)组、多柔比星(doxorubicin,DOX)2.5mg/kg组、DOX 5 mg/kg组和多巴胺(dopamine,DA)50 mg/kg组,每组30例。在肿瘤细胞接种后7d和12d,分别按相应剂量对各组实施NS、DOX或DA鼠尾静脉注射,对照组不实施任何干预。于接种后14、17、23d,分离检测,对比各组小鼠脾脏、骨髓和肿瘤中MDSC数量比例和凋亡率;比较各组肿瘤生长曲线有无差异。同期采用免疫组织化学(MaxVision)法检测对比各组瘤体、瘤旁、瘤远及瘤旁淋巴结组织中CXCL12及CXCR4表达。对各组肿瘤大小、瘤体MDSC比例与CXCL12/CXCR4表达进行相关性分析。制备肿瘤抗原特异性细胞毒性T淋巴细胞(CTL),实施荷瘤小鼠免疫治疗对比实验。荷瘤小鼠按随机数字表法分为对照组、DOX2.5mg/kg组、CTL组、DOX2.5 mg/kg+CTL组,每组30例。比较各组肿瘤生长曲线有无差异。采用SPSS22.0软件进行统计学处理。结果各用药组在肿瘤、骨髓和脾脏组织中MDSC比例均受到不同程度抑制,依次为DOX2.5组>DOX5组>DA组>对照组(P<0.05)。但各组织在不同药物刺激下MDSC凋亡率均无统计学意义(P>0.05)。各组肿瘤生长曲线比较可见DOX2.5组和其他各组间均存在统计学意义(P=0.000)。瘤体、瘤旁和淋巴组织中CXCL12和CXCR4表达较瘤远组织为高,以瘤体中表达最高。不同用药组瘤体中CXCL12和CXCR4表达均受到抑制,以DOX2.5组受抑最为显著(P=0.018,P=0.000)。各组肿瘤体积和MDSC比例�Objective To examine the effect on the expressions of CXC chemokine receptor ligand 12 (CXCL12) and CXC chemokine receptor type 4 (CXCR4) and immunotherapy of neuroblastoma (NB) by inhibiting myeloid-derived suppressor cell (MDSC) in vivo of NB-loaded BALB/c mice and explore a new method for controlling development and metastasis of NB. Methods NB-loaded BALB/c mice were established by SK-N-SH cell inoculation. The tumor specimens were removed and hematoxylin-eosin stained for observing tissue structure. Tumor-bearing mice were divided by complete randomization into normal saline (NS) injection, doxorubicin (DOX) 2.5 mg/kg injection, DOX5 mg/kg injection, dopamine (DA) 50 mg/kg injection and control groups (n=30 each). At Days 7 and 12 post-inoculation, DOX and DA were intravenously injected according to the corresponding doses while no intervention was performed in control group. The percentages and apoptotic rates of MDSC in spleen, bone marrow and tumor were detected and compared at Days 14, 17 and 23 post-inoculation. The expressions of CXCL12 and its receptor CXCR4 in tumors, peritumoral tissues, tissues distant from tumor and peripheral lymph nodes were detected and compared by immunohistochemistry (MaxVision). The volumes of tumors were compared between the groups and the correlations analyzed between volumes of tumors, ratio of MDSC and the expressions of CXCL12 and CXCR4. Then tumor antigen-specific cytotoxic T lymphocytes (CTL) were prepared. And tumor-bearing mice were divided randomly into control, DOX2.5 mg/kg, CTL and DOX2.5 mg/kg plus CTL groups (n=30 each). The tumor growth curve of each group was compared. SPSS22.0 software was utilized for statistical processing. Results All MDSC ratios were inhibited in spleen, bone marrow and tumor in different groups. The inhibitory degrees in DOX2.5 mg/kg, DOX5 mg/kg, DA50 mg/kg and control groups were in a descending order. However, no significant difference existed in apoptotic rates of MDSC between different tissues in different groups (P>0.05). Significa

关 键 词：神经母细胞瘤 肿瘤微环境 趋化因子CXCL12 受体CXCR4 小鼠 髓系抑制 性细胞 

分 类 号：R739.4[医药卫生—肿瘤]