Co-expression of herpes simplex virus thymidine kinase and Escherichia coli nitroreductase by an hTERT-driven adenovirus vector in breast cancer cells results in additive anti-tumor effects.
Co-expression of herpes simplex virus thymidine kinase and Escherichia coli nitroreductase by an hTERT-driven adenovirus vector in breast cancer cells results in additive anti-tumor effects.
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DOI：
10.3892/or.2011.1285
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发表时间：
2011-07
期刊：
影响因子：
4.2
通讯作者：
Bin Yu;Yu Zhang;Yang Zhan;Xiao Zha;Yonege Wu;Xi-zhen Zhang;Q. Dong;W. Kong;Xianghui Yu
中科院分区：
文献类型：
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作者：
Bin Yu;Yu Zhang;Yang Zhan;Xiao Zha;Yonege Wu;Xi-zhen Zhang;Q. Dong;W. Kong;Xianghui Yu
乳腺癌，特别是在晚期和转移阶段仍然难以治疗，尽管在手术技术和化疗的进步。肿瘤特异性启动子指导的自杀基因治疗和腺病毒技术可能是治疗这种晚期疾病的有前途的策略。以往的研究表明，单纯疱疹病毒胸苷激酶（HSV-TK）和更昔洛韦（GCV）与大肠杆菌硝基还原酶（Coli.NTR）和5-（azaridin-1-yl）-2，4-二硝基苯甲酰胺（CB 1954）通过重组逆转录病毒载体系统联合应用，在体外产生协同杀伤效应。我们构建了一个基于双顺反子5型腺病毒（Ad 5）的载体，该载体在人端粒酶逆转录酶（hTERT）启动子和SV 40增强子的控制下共表达疱疹病毒TK和大肠杆菌NTR。在hTERT和HSV-TK序列之后插入由内部核糖体进入位点（IRES）介导的NTR基因表达。在人乳腺癌细胞（ZR-75-30，MCF-7）中，评价了新型载体Ad-hT-TK/NTR-enh与前药联合的体内外抗肿瘤活性。结果表明，腺病毒介导的HSV-TK和NTR基因的表达与GCV和CB 1954联合应用对乳腺癌细胞具有特异性和显著的细胞毒作用。该系统的体外和体内抗肿瘤活性均优于单一自杀基因，仅略低于单独hTERT启动子驱动的HSV-TK和NTR，而腺病毒总量仅为Ad-hT-TK-enh+Ad-hT-NTR-enh的一半。这些结果表明，自杀基因HSV-TK和NTR介导的一个单一的腺病毒载体下的增强hTERT启动子的控制下的结果相加的抗肿瘤作用，并可能提供一个相对安全的策略，通过肿瘤特异性靶向治疗乳腺癌。
Breast cancers especially in the late and metastatic stages remain refractory to treatment despite advances in surgical techniques and chemotherapy. Tumor-specific promoter-directed suicide gene therapy and adenoviral technology can be promising strategies for such advanced disease. Previous studies suggested that combining herpes simplex virus thymidine kinase (HSV-TK) and ganciclovir (GCV) with Escherichia coli nitroreductase (Coli.NTR) and 5-(azaridin-1-yl)-2, 4-dinitrobenzamide (CB1954) by a recombinant retrovirus delivery system resulted in a co-operative killing effect in vitro. We constructed a bicistronic adenovirus type 5 (Ad5)-based vector which co-expresses herpes HSV-TK and Coli.NTR under the control of the human telomerase reverse transcriptase (hTERT) promoter and SV40 enhancer. NTR gene expression mediated by an internal ribosome entry site (IRES) was inserted after the hTERT and HSV-TK sequences. Anti-tumor activities of the novel vector, Ad-hT-TK/NTR-enh, combined with prodrugs were evaluated in human breast cancer cells (ZR-75-30, MCF-7) in vitro and in vivo. We showed that expression of HSV-TK and NTR genes by Ad-hT-TK/NTR-enh in combination with GCV and CB1954 resulted in specific and significant cytotoxic effects in breast cancer cells in vitro. The anti-tumor activity of this system was more efficient than that from a single suicide gene, and only slightly lower than by HSV-TK and NTR driven from separate hTERT promoters in vitro and in vivo while the total amount of adenovirus of Ad-hT-TK/NTR-enh was half that of Ad-hT-TK-enh+Ad-hT-NTR-enh. These results suggest that suicide genes HSV-TK and NTR mediated by a single adenovirus vector under the control of an enhanced hTERT promoter results in additive anti-tumor effects and may provide a relatively safe strategy for the treatment of breast cancer by tumor-specific targeting.