Adenovirus-mediated tissue-targeted expression of the HSVtk gene for the treatment of breast cancer
Adenovirus-mediated tissue-targeted expression of the HSVtk gene for the treatment of breast cancer
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DOI：
10.1038/sj.gt.3300909
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发表时间：
1999-05-01
期刊：
影响因子：
5.1
通讯作者：
Weitzman, SA
中科院分区：
文献类型：
--
作者：
Anderson, LM;Swaminathan, S;Weitzman, SA
为了开发一种治疗乳腺癌的基因疗法，我们构建了腺病毒载体，其中包含由乳腺组织特异性启动子驱动的β -半乳糖苷酶（β -gal）报告基因或单纯疱疹胸苷激酶（HSVtk）自杀基因。我们在这些载体构建中利用了人类α -乳清蛋白（hALA）基因或羊β -乳球蛋白（oBLG）基因的上游调控序列，在体外和体内将异源基因转录到乳腺癌细胞中表达。来自乳腺组织特异性报告载体的体外数据表明，与非乳腺细胞（U20S, HeLa）相比，hALA和oBLG启动子的表达确实对乳腺细胞（T47D, MCF-7, ZR75-1）具有特异性。此外，与正常的MCF-10A乳腺细胞系相比，这些载体显示出肿瘤细胞的特异性。当将这些载体全身注射到哺乳期Balb/c小鼠体内时，也显示出乳腺组织特异性，这表明这些启动子在体内腺病毒基因组的背景下保持其组织特异性表达模式。来源于T47D人乳腺癌细胞的肿瘤在裸鼠体内建立，并注射组织特异性报告细胞或自杀载体。用报告腺病毒注射肿瘤的结果表明，当T47D细胞作为已建立的肿瘤生长时，这些启动子在T47D细胞中是活跃的，我们观察到，与对照动物相比，用自杀载体注射肿瘤并系统地用更昔洛韦（150 mg/kg/天）治疗的肿瘤明显退化。此外，与对照动物相比，接受更昔洛韦联合自杀载体治疗的小鼠存活时间在35天后延长。这些数据表明，转录靶向的hALA或oBLG驱动的HSVtk基因表达可能是治疗人类乳腺癌的可行疗法。
In an effort to develop a genetic therapy for the treatment of breast cancer, we constructed adenoviral vectors containing either the beta-galactosidase (beta-gal) reporter gene or the herpes simplex thymidine kinase (HSVtk) suicide gene driven by breast tissue-specific promoters. We utilized upstream regulatory sequences from either the human alpha-lactalbumin (hALA) gene, or the ovine beta-lactoglobulin (oBLG) gene in these vector constructs to target expression of heterologous genes transcriptionally to breast cancer cells both in vitro and in vivo. Data derived from breast tissue-specific reporter vectors in vitro demonstrate that expression from the hALA and oBLG promoters are indeed specific for breast cells (T47D, MCF-7, ZR75-1) when compared with non-breast cells (U20S, HeLa). Moreover, these vectors displayed tumor cell specificity when compared with the normal MCF-10A breast cell line. These vectors also displayed breast tissue specificity when injected systemically (i.v.) into lactating Balb/c mice, which suggests that these promoters maintain their tissue-specific expression pattern within the context of the adenoviral genome in vivo. Tumors, derived from T47D human breast cancer cells, were established in nude mice and injected with either the tissue-specific reporter or suicide vectors. Results from tumors injected (i.t.) with reporter adenoviruses demonstrate that these promoters are active in T47D cells when grown as established tumors and we observed a marked regression of tumors injected with suicide vectors and treated systemically with gancyclovir (150 mg/kg/day) when compared with control animals. Moreover, mouse survival was prolonged after 35 days in mice undergoing therapy with the suicide vectors in conjunction with gancyclovir when compared with the control animals. These data suggest that the transcriptionally targeted hALA or oBLG driven expression of the HSVtk gene may be a feasible therapy for the treatment of human breast cancer.