放射性碘标记前列腺癌特异性溶瘤重组腺病毒的研究及其稳定性分析  

Study on the stability analysis and radioiodinated prostate cancer specific oncolytic recombinant adenovirus

作　　者：周家合 郝林 史振铎 贺厚光 董洋 李志刚 姜波 藏光辉 吕茜 李睿 刘颖 王洁 申友峰 何久香 韩从辉 

机构地区：[1]徐州医科大学附属徐州临床学院泌尿外科,江苏徐州221009 [2]昆山市瑞科药物研发有限公司,江苏昆山215300 [3]徐州医科大学附属徐州临床学院中心实验室,江苏徐州221009 [4]重庆威斯滕生物医药科技有限责任公司,重庆400010

出　　处：《国际泌尿系统杂志》2017年第5期660-664,共5页International Journal of Urology and Nephrology

基　　金：江苏省重点研发计划（BE2015623）;国家自然科学基金（81272557）;江苏省自然科学基金（BK2012647）;国家国际科技合作专项（2014DFA31480）;江苏六大人才高峰项目（WSW-185）;徐州市科技计划项目（XM13B079）

摘　　要：目的 核素125I标记hTERT/PSA双调控增殖型溶瘤腺病毒,构建125I-RSOAds-hTERT/PSA核素-溶瘤病毒标记物,并考察不同反应条件下病毒标记物标记率的影响.方法 125I标记采用N-溴代琥珀酰亚胺（NBS）作为氧化剂进行标记,分别设定不同浓度的溶瘤病毒和NBS进行作用,确定最佳的标记条件.分别考察125I的用量、反应时间、PH值、反应体积对125I-RSOAds-hTERT/PSA核素-溶瘤病毒标记物标记率的影响.采用凝胶柱层析法分离纯化核素-溶瘤腺病毒标记物;纸层析法测定125I-RSOAds-hTERT/PSA标记物不同时间的放射性纯度.结果 125I-RSOAds-hTERT/PSA放化纯度达95％以上,4℃冰箱放置7d其放化纯度基本稳定,保持在93％ ～94％.125I用量为0.5 μL（约0.2 mCi,7.4 MBq）,NBS用量为25μg,8×109 VP/mL,125I-RSOAds-hTERT/PSA病毒液用量为100 μL,反应时间为3 min,PH值为7.5,加入PBS体积120 μL为最优条件.结论 核素125I标记hTERT/PSA双调控增殖型溶瘤腺病毒确切可行,标记物在一定条件下放化纯度稳定.Objectives 125I is labeled to hTERT/PSA dual regulated proliferation oncolytic gland virus,and then to establish 125I-RSOAds-hTERT/PSA radionuclide oncolytic viral markers,at the same time,to investigate effects of different reaction conditions on the rate of viral markers.Methods N-bromosuccinimide （NBS） markers 125I as the oxidant,the optimal labeling conditions were determined by setting oncolytic virus and NBS with different concentrations respectively.The marker ratios of 125I-RSOAds-hTERT/PSA nuclide oncolytic viral markers in different 125I dosage,reaction time,pH value,reaction volume were observed.Radionuclide oncolytic adenovirus markers were separated and purified by by gel column chromatography.The radioactive purity of 125 I-RSOAds -hTERT/PSA markers at different time was determined by paper chromatography.Results The radioactive purity of 125I-RSOAds-hTERT/PSA markers was more than 95％,and it remained stable and remained at 93％ ～ 94％ for 7 days in the 4 degree refrigerator.In this study,125 I 0.5 μL （about 0.2 mCi,7.4 MBq）,NBS 25 μg,8 × 109 VP/mL,125l-RSOAds-hTERT/PSA virus 100 μL,reaction time 3 minutes,PH 7.5,and PBS 120 μL were the optimal reaction condition.Conclusions The method of 125I labeled hTERT/PSA double regulation of proliferation of oncolytic adenovirus is effective and feasible,and the radioactive purity of markers is stable under certain conditions.

关 键 词：前列腺肿瘤 碘放射性同位素 腺病毒科 

分 类 号：R737.25[医药卫生—肿瘤]