Ad-ING4对人膀胱癌移植瘤的生长抑制作用  

Growth inhibition of patient-derived bladder cancer by Ad-ING4

作　　者：龙建华[1] 杨慧翠[2] 杨吉成[2] 张治国[3] 侯健全[3] 欧阳骏[3] 

机构地区：[1]南华大学附属第二医院泌尿外科,湖南衡阳421001 [2]苏州大学细胞与分子生物学教研室 [3]苏州大学附属第一医院泌尿外科

出　　处：《临床泌尿外科杂志》2015年第6期541-544,548,共5页Journal of Clinical Urology

基　　金：国家自然科学基金项目(编号81070591)

摘　　要：目的:探讨Ad-ING4人膀胱癌裸鼠移植瘤的生长抑制作用及其分子机制。方法:1Ad空载体腺病毒(Ad-GFP)、Ad-ING4单基因腺病毒(Ad-ING4)分别经QBI-293A细胞感染多轮扩增后获得高效价重组病毒;2将浓度为4×107个/ml的T24膀胱癌细胞悬液在裸鼠右前肢腋下皮下注射,建立T24人膀胱癌裸鼠移植瘤模型;3将15只荷瘤裸鼠随机分为阴性对照组(PBS组)、Ad-GFP组、Ad-ING4组,3组均使用瘤体内注射干预用药,隔日1次,共6次。观测瘤体生长情况,测量肿瘤长径和短径,用公式:肿瘤体积=ab2/2(a为长径,b为短径),计算肿瘤体积,治疗3周后将裸鼠脱颈处死,摘瘤称重,用10%甲醛溶液固定标本,石蜡包埋,进行HE染色和免疫组织化学检测,并按Weidner的方法检测微血管密度(MVD)。结果:治疗后Ad-ING4组肿瘤重量为(0.93±0.06)g,分别与PBS组(1.73±0.05)g及Ad-GFP组(1.69±0.06)g相比,差异有统计学意义(P<0.05)。AdING4的抑瘤率达45%。Ad-ING4组肿瘤组织中细胞凋亡相关因子Bax和caspase-3的表达明显上调,Bcl-2的表达明显下调,肿瘤血管形成相关因子CD34表达下调,MVD降低(P<0.05)。结论:1Ad-ING4可以显著抑制膀胱癌移植瘤生长。2Ad-ING4的抑瘤机制可能和激活细胞凋亡及抑制血管形成有关。Objective：To investigate the inhibitory effects and anti-cancer mechanisms of recombinant adenovirus-mediated inhibitor of growth-4（Ad-ING4）gene on patient-derived bladder cancer in nude mice.Method：We transfected recombinant Ad-ING4 into QBI-293 Acells,after several times of amplifications,then obtained high potency contain objective gene adenovirus.Male nude mice were subcutaneously inoculated on their armpits of the right anterior limbs with 4×10^7 T24 bladder cancer cells suspension in order to construct the transplanted tumor models.Fifteen mice bearing T24 bladder cancer transplanted tumor were randomly divided into three groups：phosphate buffered saline（PBS）group,recombinant adenovirus-green fluorescent protein（Ad-GFP）group and Ad-ING4 group.The mice were intratumorally injected every other day,six times in total.Tumor progression and regression were monitored daily.Tumor volume was measured with a caliper every week and calculated by the following formula：tumor size=ab^2/2,where ais the larger and bis the smaller of the two dimensions.In addition,the tumor bearing mice were sacrificed after three-week therapy,and the tumors were removed,weighed,fixed by10% neutral formalin,and embedded in paraffin for hematoxylin and eosin staining and immunohistochemistry.We detected microvessel density（MVD）by Weidner.Result：The tumor weight of Ad-ING4 group was（0.93±0.06）g after the treatment,compared with PBS group（1.73±0.05）g and Ad-GFP（1.69±0.06）g,respectively.The difference was statistically significant（P〈0.05）.The ratio of tumor suppression of Ad-ING4 group was45%.In Ad-ING4 group the expressions of Bax and caspase-3were up-regulated,and Bcl-2was down-regulated.Moreover,CD34 which correlated with angiopoiesis was down-regulated and MVD decreased（P〈0.05）.Conclusion：Ad-ING4 can noticeably inhibit the growth of transplanted tumor.The anti-cancer molecular mechanisms of Ad-ING4 may be associated with activating apoptosis and inhibition of angiopoiesis.

关 键 词：ING4 T24人膀胱癌细胞 移植瘤 基因治疗 

分 类 号：R737.14[医药卫生—肿瘤]