融合基因重组卡介苗对肿瘤的抑制作用  

Inhibiting effect function of rBCG using fusion gene on tumors

作　　者：闫迎春[1] 王子豪 李运清[1] 杜峰[1] 刘昂 陈廷[1] 薛庆节[1] YAN Yingchun;WANG Zihao;LI Yunqing;DU Feng;LIU Ang;CHEN Ting;XUE Qingjie(Jining Medical University,Jining,Shandong 272067,China)

机构地区：[1]济宁医学院,山东济宁272067

出　　处：《中国热带医学》2020年第2期102-105,111,共5页China Tropical Medicine

基　　金：国家自然科学基金项目(No.31500056);山东省重点研发计划项目(No.2018GSF118137);山东省自然科学基金项目(No.ZR2012HM037);山东省医药卫生科技发展计划项目(No.2017WS339);山东省高等学校科技计划项目(No.J17KB085);济宁医学院青年教师科研扶持基金(No.JY2017KJ019);济宁医学院国家自然科学基金培育项目资助(2016);济宁市科技助推新旧动能转换计划项目(No.2017SMNS001);2018年国家级大学生创新创业训练计划项目(No.201810443034).

摘　　要：目的研究EB病毒基因BZLF1和LMP2融合基因重组卡介苗(recombinant BCG,rBCG)对肿瘤的抑制作用。方法建立EB病毒阳性肿瘤(NPRC18)荷瘤小鼠(C57BL/6J)模型,根据注射的药物不同,分为rBCG组、BCG组、转pMV261载体BCG组(pMV261+BCG)、PBS组。采取颈背部皮下多点注射方式进行免疫,对荷瘤小鼠肿瘤重量、成瘤时间、存活情况进行分析,C57BL/6J小鼠肿瘤抑瘤实验结束后,将小鼠的肿瘤裂解后用流式检测CD4+T、CD8+T细胞增殖情况,HE染色分析小鼠肿瘤组织中淋巴细胞浸润情况;使用统计学软件SPSS 19.0对rBCG抑瘤效果进行单因素方差分析(One-way ANOVA)。结果重组BCG组分别用BZLF1和LMP2一抗杂交后,出现杂交目的蛋白,其蛋白质分子量约为70 KD,与对照相比,重组卡介苗对肿瘤抑制作用明显,小鼠成瘤时间延迟,平均成瘤时间约22 d,与对照PBS注射组8 d相比,其差异均有统计学意义(P≤0.01);PBS组肿瘤重量为6.8 g,重组BCG组肿瘤重量为1.01 g,抑瘤率约为81%。经流式分析,重组BCG组产生的CD4+T、CD8+T细胞高于对照组。与对照组相比,rBCG延长了小鼠的生存时间,延缓了肿瘤生长和形成时间。结论重组卡介苗在小鼠体内对肿瘤细胞具有抑制作用。Objective The inhibition on tumor of EB virus BZLF1 and LMP2 fusion gene recombinant BCG vaccine(recombinant BCG,rBCG)were researched.Methods EB virus-positive tumor cells(NPRC18)cancer models were established with C57 BL/6 J mice.According to the injection of drugs,the C57 BL/6 J micewere divided into rBCG group,BCG group,pMV261+BCG group,PBS group.The tumor weight,time of tumor formation and survival of tumor-bearing were analyzed after subcutaneous multi-point injection immunization in backand neck of the mice.After the tumor inhibition experiment of C57 BL/6 J mice,the tumor cells were cleaved,and the proliferation of CD4+T and CD8+T cells was detected by flow cytometry.HE staining was used todetect and analyze lyphocyte infiltration.Single factor analysis method of variance(One-way ANOVA)was used for evaluation of rBCG’s inhibition by SPSS 19.0 statistical software.Results Recombinant BCG group have a hybrid with BZLF1 and LMP2 respectively,target protein molecular weight was about 70 KD,compared with controls,the recombinant BCG vaccine has an obvious effect of tumor suppression,time delay in the formation of the tumor in mice,the average time of the tumor formation time is about 22 days,compared with the control PBS injection group 8 days,the difference was significance(P≤0.01).The tumor weight of the PBS group was 6.8 g,the recombinant BCG group was 1.01 g,the tumor inhibition rate was about 81%.By flow cytometry analysis,CD4+T and CD8+T cells were significantly higher in the recombinant BCG group than in the control group.Compared with the control group,rBCG prolonged survival time of mice,slowed tumor growth and delayed tumor formation time significantly.Conclusion rBCG can inhibit tumor cells growing in mice.

关 键 词：重组卡介苗 EB病毒阳性肿瘤 融合基因 

分 类 号：R730.51[医药卫生—肿瘤]