Targeted gene therapy of ovarian cancer using an ovarian-specific promoter.
Targeted gene therapy of ovarian cancer using an ovarian-specific promoter.
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DOI：
10.1006/gyno.2001.6490
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发表时间：
2002-02
影响因子：
4.7
通讯作者：
R. Bao;M. Selvakumaran;T. C. Hamilton
中科院分区：
文献类型：
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作者：
R. Bao;M. Selvakumaran;T. C. Hamilton
目的 使用巨细胞病毒等启动子进行癌症“自杀”基因治疗，由于缺乏前体药物激活的特异性，可能会对正常组织造成严重的毒性。因此，我们研究了卵巢癌的基因治疗，使用卵巢特异性启动子（OSP1）来限制卵巢癌细胞前体药物激活酶HSVtk的合成。方法构建HSVtk表达质粒pOSP1-HSVtk并转染至卵巢癌细胞系OVCAR3中。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑法评价稳定转染子的更昔洛韦(GCV)敏感性。通过比较卵巢癌细胞系和非卵巢癌细胞系转染 pOSP1-HSVtk 后对 GCV 的敏感性来评估该启动子的组织特异性。将一种对 GCV 敏感的转染子植入免疫受损小鼠腹膜内，随后用 GCV 治疗小鼠。此外，该卵巢癌生存模型用于评估阳离子脂质介导的 pOSP1-HSVtk 基因递送随后进行 GCV 治疗的体内功效。结果 与亲本细胞系和载体转染的 OVCAR3 细胞系相比，携带 OSP1-HSVtk 的 OVCAR3 细胞的稳定转染子对 GCV 处理更加敏感。 OSP1-HSVtk 可以特异性地使 OVCAR3 卵巢癌细胞系对 GCV 敏感。移植 OVCAR3 转染子并接受 GCV 治疗的 SCID 小鼠比未接受 GCV 治疗的小鼠存活时间更长 (P = 0.032)。在 OVCAR3 生存模型中，由阳离子脂质 (GL67) 介导的体内基因传递和 GCV 治疗产生了更长的生存期 (P = 0.016)。结论 OSP1启动子可以选择性地指导卵巢癌的自杀基因治疗，并且与直接注射质粒相比，使用阳离子脂质GL67作为递送载体可以提高体内疗效。
OBJECTIVES The "suicide" gene therapy of cancer using promoters such as cytomegalovirus could cause severe toxicity to normal tissues due to a lack of specificity of prodrug activation. Therefore, we investigated gene therapy of ovarian cancer using ovarian-specific promoter (OSP1) to limit the synthesis of the prodrug activating enzyme HSVtk to ovarian cancer cells. METHODS The HSVtk expressing plasmid pOSP1-HSVtk was created and transfected into an ovarian cancer cell line OVCAR3. The ganciclovir (GCV) sensitivity of the stable transfectants was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Tissue specificity of this promoter was evaluated by comparing the sensitivity to GCV between ovarian and nonovarian cancer cell lines after they were transfected with pOSP1-HSVtk. One transfectant sensitive to GCV was implanted intraperitoneally to immunocompromised mice which were treated subsequently with GCV. Furthermore, this ovarian cancer survival model was used to evaluate the in vivo efficacy of cationic lipid mediated pOSP1-HSVtk gene delivery followed by GCV treatment. RESULTS Stable transfectants of OVCAR3 cells bearing OSP1-HSVtk became more sensitive to GCV treatment compared to the parental cell line and vector transfected OVCAR3 cell line. OSP1-HSVtk could specifically sensitize the OVCAR3 ovarian cancer cell line to GCV. SCID mice transplanted with the OVCAR3 transfectant and treated with GCV survived longer than the mice without GCV treatment (P = 0.032). In vivo gene delivery mediated by a cationic lipid (GL67) followed by GCV treatment yielded a longer survival in the OVCAR3 survival model (P = 0.016). CONCLUSIONS The OSP1 promoter can selectively direct suicide gene therapy of ovarian cancer and the in vivo efficacy is improved by using a cationic lipid GL67 as delivery vehicle as opposed to the direct injection of plasmid.