选择性增殖腺病毒CNHK500治疗乳腺癌的实验研究  

A Conditionally Replicating Adenovirus CNHK500 for the Treatment of Breast Cancer

作　　者：李月敏[1] 宋三泰[1] 江泽飞[1] 徐建明[1] 张琪[2] 李明颖[1] 钱炎珍 崔贞福[2] 钱其军[2] 

机构地区：[1]军事医学科学院附307医院肿瘤四科,北京100039 [2]第二军医大学东方肝胆外科医院,上海200438 [3]上海新霁生物新科技有限公司,上海201203

出　　处：《中国肿瘤生物治疗杂志》2005年第2期124-128,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家863计划重点资助项目(2001AA217031)国家自然科学基金国际合作项目(30120160824)

摘　　要：目的:观察选择性增殖腺病毒CNHK500对乳腺癌的特异性杀伤作用。方法:行病毒增殖实验和细胞生长抑制实验,验证CNHK500选择性复制和杀伤能力;Westernblot检测腺病毒E1A和E1B在细胞中的表达。结果:CNHK500在乳腺癌细胞中复制能力与野生型腺病毒wtAd5相似,较ONYX-015增殖能力强。在正常成纤维细胞中CNHK500病毒增殖能力明显减弱,较wtAd5增殖能力弱1000倍左右。CNHK500可有效杀伤乳腺癌细胞株;而CNHK500对正常成纤维细胞的杀伤力较wtAd5减弱约100倍。CNHK500病毒的E1A可以选择性在端粒酶阳性的乳腺癌细胞株中表达,在端粒酶阴性的正常成纤维细胞株BJ中不表达,CNHK500可以选择性地在缺氧条件下表达E1B。动物实验结果显示,静脉注射CNHK500可以显著抑制MCF-7乳腺癌细胞裸鼠移植瘤的生长,治疗效果与给药剂量相关。结论:肿瘤选择性增殖腺病毒CNHK500可选择性在端粒酶阳性的乳腺癌细胞中复制,并产生体内外杀伤作用。Objective:To evaluate the selectively oncolytic effect of conditionally replicating adenovirus CNHK500 in breast cancer. Methods:We used virus proliferation assay cell viability assay to evaluate the proliferation and cytolysis selectivity of CNHK500. And we used Western-blot to confirm the expression of adenovirus CNHK500 E1A and E1B in cancer and normal cells. Results:The CNHK500 virus proliferation ability in breast cancer cell lines is similar to that of wtAd5, better than that of ONYX-015 virus. However, CNHK500 virus replicate 1000-fold less than that of wtAd5 in normal fibroblast cell lines. CNHK500 can effectively kill breast cancer cell, while it shows attenuated cytolysis in normal fi-broblast cells, with about 100-fold less than that of wtAd5. CNHK500 E1A is expressed in telomerase-positive breast cancer cells but not in lelomerase-negative normal fibroblast cells. E1B protein can be detected under hypoxia condition but not in normoxia conditions. Intravenous injections of CNHK500, yielded significant tumor growth delay in the telomer-ase-positive breast cancer xenografts though the MCF-7 represent a very fast growing tumor cell line. Antitumor efficacy of replication-competent adenovirus CNHK500 in vivo was associated with increased dosage of CNHK500. Conclusions:The results prove that CNHK500 has highly proliferation selectivity and potent cytolysis effect on breast cancer.

关 键 词：肿瘤增殖病毒 乳腺癌 基因治疗 端粒酶 HRE 

分 类 号：R737.9[医药卫生—肿瘤]