β-arrestin1过表达抑制Molt-4细胞异种移植小鼠急性T淋巴细胞白血病进展  

β-arrestin1 overexpression suppresses progression of human T-cell acute lymphatic leukemia Molt-4 cell xenograft in mice

作　　者：张佳[1] 舒逸[1] 张虹洋 蒋婷婷 宫茂源 朱丹 王皓飚 邹琳[1] ZHANG Jia;SHU Yi;ZHANG Hongyang;JIANG Tingting;GONG Maoyuan;ZHU Dan;WANG Haobiao;ZOU Lin(Center for Clinical Molecular Medicine,Children's Hospital,Ministry of Education Key Laboratory of Child Development and Disorders,National Clinical Research Center for Child Health and Disorders,China International Science and Technology Cooperation Base of Child Development and Critical Disorders,Children's Hospital of Chongqing Medical University,Chongqing Engineering Research Center of Stem Cell Therapy,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院临床分子医学中心//儿童发育疾病研究教育部重点实验室//国家儿童健康与疾病临床医学研究中心//儿童发育重大疾病国家国际科技合作基地//重庆市干细胞治疗工程技术研究中心,重庆400014

出　　处：《南方医科大学学报》2020年第5期654-660,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(81570142,81870126)。

摘　　要：目的探讨β-arrestin1对Molt-4细胞异种移植急性T淋巴细胞白血病(T-ALL)NCG小鼠模型疾病进展的影响。方法建立稳定表达荧光素酶的人Molt-4细胞系,酶标仪检测其荧光素酶强度;构建稳定过表达或敲低β-arrestin1和空载体的Molt-4细胞系,设为β1组、Siβ1组和Scram组并分别接种于亚致死计量辐射后NCG小鼠体内;记录小鼠移植后体质量及生存时间;利用活体荧光成像系统检测移植小鼠模型体内肿瘤进展;移植后16 d处死小鼠,取肝和脾做切片染色,检测肿瘤细胞浸润。结果成功构建稳定表达荧光素酶Molt-4细胞系(Molt4-Fluc)。在Molt4-Fluc基础上成功构建过表达β-arrestin1(β1)、敲低β-arrestin1(Siβ1)或空载体(Scram)的Molt-4细胞系。NCG小鼠T-ALL异种移植模型显示,β1组、Siβ1组及Scram组Molt4-Fluc细胞在受体NCG小鼠中进展,导致NCG小鼠在移植后体质量降低。小鼠生存时间显示,β1组细胞受体小鼠生存时间(43 d)显著长于Scram组细胞受体小鼠(33 d),P<0.001;Siβ1组细胞受体小鼠生存时间(20 d)显著短于Scram组细胞受体小鼠(33 d),P<0.001;活体荧光成像结果显示,β1组细胞受体小鼠体内T-ALL细胞荧光强度显著弱于Scram组细胞受体小鼠,Siβ1组细胞受体小鼠体内T-ALL细胞荧光强度强于Scram组细胞受体小鼠;处死移植后16 d小鼠,组织染色结果显示,β1组细胞受体小鼠肝、脾内肿瘤细胞浸润较Scram组细胞受体小鼠少。结论β-arrestin1过表达可抑制Molt-4细胞异种移植后小鼠T-ALL进展。Objective To investigate the effect of β-arrestin1 overexpression on tumor progression in a NCG mouse model bearing T-cell acute lymphocytic leukemia(T-ALL)Molt-4 cell xenograft.Methods Molt-4 cells were tagged with firefly-luciferase(F-Luc)by lentiviral infection,and fluorescence intensity of the cells was detected using a luminescence detector.Molt-4 cell lines with β-arrestin1 overexpression or knockdown were constructed by lentivirus infection and injected via the tail vein in sub-lethal irradiated NCG mice.Body weight changes and survival time of the xenografted mice were observed,and the progression of T-ALL in the mice was evaluated using an in vivo fluorescence imaging system.Sixteen days after xenografting,the mice were euthanatized and tumor cell infiltration was observed in the slices of the liver and spleen.Results We successfully tagged Molt-4 cells with F-Luc and overexpressed or knocked down β-arrestin1 in the tagged cells.Bioluminescent imaging showed obvious luminescence catalyzed by F-Luc in Molt-4 cells.After injection of Molt-4-Luc cells into irradiated NCG mice,a gradual enhancement of luminescence in the xenografted mice was observed over time,while the body weight of the mice decreased.Compared with the control mice,the mice xenografted with β-arrestin1-overexpressing Molt-4 cells had significantly prolonged survival time(P<0.001),while the survival time of the mice xenografted with Molt-4 cells with β-arrestin1 knockdown was significantly shortened(P<0.001).Histological examination revealed fewer infiltrating tumor cells in the liver and spleen of the mice xenografted with β-arrestin1-overexpressing Molt-4 cells in comparison with the mice bearing parental Molt-4 cell xenografts.Conclusions β-arrestin1 overexpression suppresses tumor progression in mice bearing Molt-4 cell xenograft.

关 键 词：T-ALL β-arrestin1 小鼠异种移植模型 活体荧光成像 MOLT-4细胞 

分 类 号：R733.71[医药卫生—肿瘤]