腺病毒介导的色素上皮衍生因子对小鼠黑色素瘤肺转移的抑制作用  

Inhibitive effect of recombinant adenoviruses loaded with pigment epithelium-derived factor gene on pulmonary metastases of melanoma in mice

作　　者：陈巧玲[1] 白亦光[2] 付曦[1] 何朗[1] 冯刚[3] CHEN Qiaoling BAI Yiguang FU Xi HE Lang FENG Gang(Department of Oncology, Nanchong Central Hospital, Second Clini- cal College of North Sichuan Medical College, Nanchong 637000, China Department of Orthopedics, Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College Research Institute of Tissue Engineering and Stem Cell,Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College)

机构地区：[1]川北医学院第二临床医学院,南充市中心医院肿瘤科,南充637000 [2]川北医学院第二临床医学院、南充市中心医院骨科 [3]川北医学院第二临床医学院、南充市中心医院组织工程与干细胞研究所

出　　处：《山西医科大学学报》2017年第6期534-538,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30872614);四川省教育厅理科重点项目(15ZA0216,15ZB0201);南充市科技局科技支撑项目(14A0017,14A0022);川北医学院科研发展计划资助项目(CBY14-A-ZD02,CBY15-A-ZD02);四川省卫计委科研基金资助项目(16PJ202)

摘　　要：目的探讨腺病毒介导的色素上皮衍生因子(Ad-PEDF)对小鼠B16F10黑色素瘤肺转移模型的治疗作用。方法在体外用MTT实验测定Ad-PEDF感染复数(multiplicity of infection,MOI)为50,100,150,200,250时对B16F10细胞增殖的抑制作用,Transwell实验检测Ad-PEDF对B16F10细胞侵袭的抑制作用,以PBS和空载腺病毒(Ad-null)作为对照。在体内通过尾静脉接种小鼠B16F10细胞建立肺转移模型,将小鼠随机分为3组,分别经尾静脉注射100μl PBS、100μl Ad-null(1×108PFU)和100μl Ad-PEDF(1×108PFU)。隔日注射1次,共5次。观察静脉注射Ad-PEDF对黑色素瘤肺转移结节数量以及肺湿重的影响。对肺转移灶进行TUNEL染色检测肿瘤细胞的凋亡情况。结果 MTT实验显示,Ad-PEDF在不同的MOI值下均能对B16F10细胞产生作用,并随着MOI值的增加抑制率逐渐升高,与Ad-null组相比,差异有统计学意义(P<0.01)。Transwell实验显示,AdPEDF组与PBS组和Ad-null组相比,穿膜的肿瘤细胞数明显下降,侵袭能力受到显著抑制(123.3±21.9 vs 220.3±26.8,213.5±35.5,P<0.01)。体内实验结果提示,Ad-PEDF组小鼠的肺部转移灶较PBS组和Ad-null组明显减少(47.7±14.1 vs 75.1±20.9,73.3±17.3,P<0.05)。而Ad-PEDF组的小鼠肺湿重[(0.26±0.06)g]也明显低于PBS组[(0.39±0.05)g]和Ad-null组[(0.37±0.07)g],差异有统计学意义(均P<0.05)。TUNEL实验显示,Ad-PEDF组肺转移灶肿瘤细胞凋亡率较PBS组和Ad-null组增多(25.3%±3.3%vs 3.9%±1.2%,4.1%±1.4%,P<0.01)。结论腺病毒介导的PEDF能够显著抑制小鼠黑色素瘤肺转移灶的形成,其作用机制可能为抑制肿瘤细胞增殖和侵袭,以及诱导肿瘤细胞凋亡。Objective To explore the inhibitive effect of recombinant adenoviruses loaded with pigment epithelium-derived factor gene on pulmonary metastases of melanoma B16F10 in mice. Methods MTY and Transwell assays were performed to observe the inhibitive effect of Ad-PEDF on B16F10 cells proliferation and invasion ability of B16F10 cells with PBS and adenovirus vector（Ad-null） as the controls. The C57BL/6 mice pulmonary metastases models of melanoma were established by the injection of B16F10 ceils via tail vein. Then 30 tumor-bearing mice were randomly divided into 3 groups：PBS group, Ad-null group and Ad-PEDF group. The mice were intra- venously administrated with PBS（ 100 μ1） ,adenoviruses vector（ 1 ×10^8 PFU, 100μl） or Ad-PEDF（ 1 ×10s PFU, 100 μl） every other day for five times, respectively. The influence of Ad-PEDF on the pulmonary metastases nodule numbers was observed, and the wet weight of the lungs in mice was calculated. The apoptosis of the tumor cells in metastasis nodules was detected by TUNEL assay. Re- sults MrlT assay showed that B16F10 cells proliferation decreased in a dose-dependent manner after treatment with different multiplici- ty of infection（MOI） of Ad-PEDF（ P 〈 0. 01 ）. Transwell assay demonstrated that the number of B16F10 cells migrating to the other side of the membrane in Ad-PEDF group was markedly decreased compared with PBS group or Ad-null group （ 123.3 ± 21.9 vs 220. 3 ± 26. 8,213.5 ± 35.5 ,P 〈 0. 01 ）. The in vivo results showed that the number of pulmonary metastases nodules in Ad-PEDF group was less than in PBS group and Ad-null group（47.7 ± 14. 1 vs 75. 1±20. 9,73.3 ± 17.3 ,P 〈0. 05）. And the wet weight of the lungs was reduced in Ad-PEDF group compared with PBS group and Ad-null group [ （0. 26 ± 0. 06） g vs （0. 39 ± 0. 05 ） g, （0. 37 ± 0.07 ） g, P 〈 0. 05 ]. The TUNEL assay showed that Ad-PEDF increased the apoptosis rate of tumor cells in lung metastasis foci compared with PBS and Ad-null（25.3%±3.3% vs 3.9%±1

关 键 词：色素上皮衍生因子 腺病毒载体 黑色素瘤 肺转移 

分 类 号：R739.5[医药卫生—肿瘤]