加载MAGE-A10/hβD-2双基因溶瘤病毒对膀胱癌细胞增殖及肿瘤生长的影响  

Effect of oncolytic virus loaded with MAGE-A10/hβD-2 double gene on bladder cancer cell proliferation and tumor growth

作　　者：张建军 蔡龙俊 蔡维奇 ZHANG Jianjun;CAI Longjun;CAI Weiqi(Kewen College,Jiangsu Normal University,Jiangsu Province,Xuzhou 221132,China;Department of Urology,the Affiliated Suqian Hospital of Xuzhou Medical University,Suqian Hospital of Nanjing Drum-Tower Hospital Group,Jiangsu Province,Suqian 223800,China)

机构地区：[1]江苏师范大学科文学院,江苏徐州221132 [2]徐州医科大学附属宿迁医院暨南京鼓楼医院集团宿迁医院泌尿外科,江苏宿迁223800

出　　处：《中国医药导报》2022年第9期21-26,45,共7页China Medical Herald

基　　金：江苏省高等学校自然科学研究面上项目(19KJB320008);宿迁市市级指导性科技计划项目(Z2020106)。

摘　　要：目的探讨加载黑色素瘤相关抗原A10(MAGE-A10)和人β防御素2(hβD-2)基因的溶瘤病毒SG502-MAGE-A10-hβD-2对膀胱癌增殖及肿瘤生长的作用。方法构建加载MAGE-A10和hβD-2基因的溶瘤病毒SG502-MAGE-A10-hβD-2,半数组织培养感染剂量法测定病毒滴度。CCK-8法测定溶瘤病毒SG502及SG502-MAGE-A10-hβD-2对T24及MB49细胞系增殖的影响,其中SG502组加入SG502病毒,SG502-MAGE-A10-hβD-2组加入SG502-MAGE-A10-hβD-2病毒,对照组加入PBS,分别培养24、48、72、96 h。采用MB49细胞构建C57BL/6小鼠皮下移植瘤模型,待肿瘤生长至100~150 mm^(3)后将成瘤的18只小鼠按照随机数字表法分成对照组、SG502组及SG502-MAGE-A10-hβD-2组,每组各6只。对照组给予50μl生理盐水进行瘤内多点注射,SG502组给予含有2×10^(8)个病毒颗粒数的SG502病毒的50μl生理盐水溶液进行瘤内多点注射,SG502-MAGE-A10-hβD-2组给予含有2×10^(8)个病毒颗粒数的SG502-MAGE-A10-hβD-2病毒的50μl生理盐水溶液进行瘤内多点注射,每周2次。瘤内注射5周后检测各组小鼠肿瘤的体积、成瘤组织MAGE-A10及hβD-2蛋白的表达、Ki67的表达及血清中TNF-α、IFN-γ的含量。结果成功构建溶瘤病毒SG502-MAGE-A10-hβD-2,纯化后病毒滴度为2.05×10^(10) PFU/ml。SG502组、SG502-MAGE-A10-hβD-2组组内各时间点的T24及MB49细胞抑制率比较,差异均有统计学意义(P<0.05),SG502组与对照组24、48、72、96 h的T24及MB49细胞抑制率比较,差异均有统计学意义(P<0.05),SG502-MAGE-A10-hβD-2组与SG502组24、48、72、96 h的T24及MB49细胞抑制率比较,差异均有统计学意义(P<0.05)。SG502组、SG502-MAGE-A10-hβD-2组组内各时间点的肿瘤体积比较,差异均有统计学意义(P<0.05),SG502组与对照组14、21、28、35、42 d的肿瘤体积比较,差异均有统计学意义(P<0.05),SG502-MAGE-A10-hβD-2组与SG502组14、21、28、35、42 d的肿瘤体积比较,差异均有统计学意义(P<0.05)。SG502组肿瘤重量均小于�Objective To explore the effect of oncolytic virus SG502-MAGE-A10-hβD-2 loaded with melanoma-associated antigen A10(MAGE-A10)and human beta defensin 2(hβD-2)genes on the proliferation and tumor growth of bladder cancer cell lines.Methods The oncolytic virus SG502-MAGE-A10-hβD-2 loaded with MAGE-A10 and hβD-2 genes was constructed.Virus titer was determined by 50% tissue culture infectious dose.CCK-8 method were used to determine the effects of oncolytic viruses SG502 and SG502-MAGE-A10-hβD-2 on the proliferation of T24 and MB49 cells.SG502 group was added with SG502 virus,SG502-MAGE-A10-hβD-2 group was added with SG502-MAGE-A10-hβD-2 virus,and the control group was added with PBS for 24,48,72,96 h,respectively.The MB49 cell was used to construct a C57BL/6 mouse subcutaneously transplanted tumor model.After the tumor grew to 100-150 mm^(3),the eighteen mice were divided into control group,SG502 group and SG502-MAGE-A10-HβD-2 group according to the random number table method,with six mice in each group.The control group was given 50μl normal saline for intratumoral multipoint injection.SG502 group was given 50μl normal saline solution containing SG502 virus with 2×10^(8) virus particles for intratumoral multipoint injection.SG502-MAGE-A10-hβD-2 group was given 50μl normal saline solution containing SG502-MAGE-A10-hβD-2 virus particles of 2×10^(8) in intratumoral multipoint injection,two times a week.The tumor volume,the expression of MAGE-A10 and hβD-2 proteins in tumor-forming tissues,the expression of Ki67 and the contents of TNF-α and IFN-γ in serum were detected five weeks after intratumoral injection.Results The oncolytic virus SG502-MAGE-A10-hβD-2 was successfully constructed,and the titer of the purified virus was 2.05×10^(10) PFU/ml.The inhibition rates of T24 and MB49 cells at each time point in SG502 group and SG502-MAGE-A10-hβD-2 group were statistically significant(P<0.05).There were statistically significant differences in T24 and MB49 cell inhibition rates between SG502 group and

关 键 词：膀胱癌 溶瘤病毒 细胞增殖 基因治疗 

分 类 号：R737.14[医药卫生—肿瘤]