尾静脉注射及皮下接种B-LCL细胞建立EB病毒相关淋巴瘤动物模型的比较研究  

Establishment of mouse models of lymphoma with dual-labeled EBV-immortalized lymphoblastoid cell lines by intravenous versus subcutaneous injection

作　　者：方兰兰 董廷 周颖 孙语璐 高阳 熊云青 顾潮江 Lanlan Fang;Ting Dong;Ying Zhou;Yulu Sun;Yang Gao;Yunqing Xiong;Chaojiang Gu(The College of Life Sciences and Health,Wuhan University of Science and Technology,Wuhan 430065,China)

机构地区：[1]武汉科技大学生命科学与健康学院,武汉市430065

出　　处：《中国肿瘤临床》2023年第24期1243-1247,共5页Chinese Journal of Clinical Oncology

基　　金：国家重点研发计划“政府间/港澳台重点专项”项目(编号:2017YFE0129100);湖北省重点研发计划项目(编号:2023BCB016)资助。

摘　　要：目的:构建绿色荧光蛋白(green fluorescent protein,GFP)和荧光素酶(luciferase,Luc)双标记的EB病毒(Epstein-Barr virus,EBV)感染的人B淋巴母细胞系(B lymphoblastoid cell lines,B-LCL)应用于肿瘤模型,并比较尾静脉注射和皮下注射人B-LCL细胞建立小鼠模型的优缺点。方法:通过二代慢病毒转染、嘌呤霉素筛选构建GFP/Luc双标的B-LCL细胞系(B lymphoblastoid cell lines double-tagged with GFP and Luc,B-LCL-GL),接种低、中和高3种剂量的细胞至NPG[(NOD)/Prkdcscid/IL-2Rγ^(null)]小鼠皮下或尾静脉内,建立皮下移植瘤模型与血行性转移瘤模型并成像分析。结果:B-LCL-GL细胞中的GFP阳性细胞比例为92.5%,荧光素酶平均发光强度为4.80E+08 Photons/s,远高于B-LCL组。在血行性转移瘤模型中,连续成像结果表明从第7天到第28天随着肿瘤移植时间的延长,肿瘤最先定植在腹腔后又转移到全身。在皮下移植瘤模型中,第7天时3组小鼠均可以检测到肿瘤细胞在小鼠皮下接种处发出了中心强、周围弱的荧光信号,第28天时高剂量组肿瘤转移到全身。相同接种剂量分别通过尾静脉和皮下接种时,肿瘤生物发光均无显著性差异(P>0.05);小鼠存活情况显示低、中、高各剂量尾静脉注射组在第100天时全部死亡,低剂量和中剂量皮下注射组小鼠带瘤生存长达100天且未出现小鼠死亡,可允许更长时间进行实验。结论:成功构建GFP/Luc双标的B-LCL细胞系,尾静脉注射和皮下注射异种移植B-LCL细胞两种方法均可成功建立EBV-LCLs肿瘤模型,并可通过成像示踪定位和精准定量肿瘤大小,为肿瘤特性研究及抗肿瘤药物筛选提供了研究平台。Objective:To establish a green fluorescent protein(GFP)and firefly luciferase(Luc)double-labeled Epstein-Barr virus(EBV)infected B lymphoblastoid cell lines(B-LCL)and apply them to mouse models,then compare the advantages and disadvantages of models inoculated by intravenous(IV)or subcutaneous(SC).Methods:B lymphoblastoid cell lines double-tagged with GFP/Luc(B-LCL-GL)were constructed through lentivirus transduction,puromycin intervention.Subcutaneous xenograft and hematogenous metastasis models were respectively established by subcutaneous or intravenous injection of B-LCL-GL cells at three concentrations in(NOD)/Prkdcscid/IL-2Rγ^(null)(NPG)mice for in vivo bioluminescence imaging.Results:In the B-LCL-GL group,the ratio of the GFP-positive cell population was 92.5%,and the average luminescence intensity was as high as 4.80E+08 Photons/s,which was considerably higher than that of untreated B-LCLs.In the hematogenous metastasis models,tumor bioluminescence was initially located in the peritoneal area and then spread throughout the entire body between 7 and 28 days.In the subcutaneous xenograft models,strong central and weak peripheral tumor-related bioluminescence signal was detected on day 7 in the three groups,which then spread throughout the body on day 28 in the high-dose group.Taken together,there was no significant difference in tumor progression between the two routes of administration when using the same dose of BLCL-GL cells.However,the survival analysis indicated that the IV injection group,in which all the mice ultimately died,had a shorter time frame for testing than that of the SC injection group,in which the mice survived until day 100 in the low-dose and medium-dose groups,thus allowing for long-term testing.Conclusions:GFP and Luc dual-positive B-LCLs were successfully established to generate hematogenous metastasis and subcutaneous xenograft models,which allow the monitoring of the location and size of lymphomas in vivo.It provide platform for the study of tumor characteristics and selecting anti

关 键 词：B细胞淋巴瘤 EB病毒 GFP/Luc 双标记疾病模型 静脉注射 皮下注射 

分 类 号：R733.1[医药卫生—肿瘤] R-332[医药卫生—临床医学]