Strategies to accomplish targeted expression of transgenes in ovarian cancer for molecular therapeutic applications.
Strategies to accomplish targeted expression of transgenes in ovarian cancer for molecular therapeutic applications.
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发表时间：
2001-08
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通讯作者：
Enrique Casado;J. Gómez-Navarro;Masato Yamamoto;Y. Adachi;C. Coolidge;W. Arafat;S. Barker;Minghui Wang;P. Mahasreshti;A. Hemminki;Manuel González-Barón;Manuel González-Barón;M. Barnes;T. Pustilnik;G. Siegal;G. Siegal;R. Alvarez;David T. Curiel
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作者：
Enrique Casado;J. Gómez-Navarro;Masato Yamamoto;Y. Adachi;C. Coolidge;W. Arafat;S. Barker;Minghui Wang;P. Mahasreshti;A. Hemminki;Manuel González-Barón;Manuel González-Barón;M. Barnes;T. Pustilnik;G. Siegal;G. Siegal;R. Alvarez;David T. Curiel
目的探讨中期因子(MK)和环氧合酶-2(COX-2)基因启动子区作为肿瘤特异性启动子在卵巢癌靶向基因治疗中的作用。已建立的实验设计通过腺病毒载体转导卵巢癌和间皮细胞，腺病毒载体含有在MK、COX-2或巨细胞病毒(CMV)启动子控制下表达的报告基因或胸苷激酶基因。给SCID或C57BL/6小鼠腹腔注射。用这些相同的载体。用荧光素酶和四甲基偶氮唑盐(3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium)比色法分别检测报告基因的表达和细胞毒作用。通过组织学评价进行体内急性毒性评价。结果除原代卵巢癌细胞外，所有卵巢癌细胞系中均有MK和COX-2启动子的一致激活。相反，报告显示，用含有测试启动子的腺病毒转导的间皮细胞的报告活性降低，这对COX-2启动子尤其明显。此外，在体内实验中，COX-2启动子在肝脏和腹膜中的报告基因水平显著低于对照启动子。Adcox-2 MTK对肿瘤细胞的杀伤作用与AdCMVTK相当。然而，与对照组相比，观察到明显的不同毒性模式，有利于Adcox-2 MTK治疗的动物。结论这些数据清楚地表明，COX-2启动子提供的转录调控是肿瘤特异性的，能够减轻正常组织的相关毒性，同时在卵巢癌分子化疗方法的背景下保持治疗效果。
PURPOSE The purpose of the study was to determine the capability of the midkine (MK) and cycooxygenase-2 (cox-2) gene promoter regions to function as tumor-specific promoters for use in targeted gene therapy of ovarian cancer. EXPERIMENTAL DESIGN Established and primary ovarian cancer and mesothelial cells were transduced by adenoviral vectors containing a reporter or thymidine kinase gene expressed under the control of the MK, cox-2, or cytomegalovirus (CMV) promoters. SCID or C57BL/6 mice were injected i.p. with these same vectors. In vitro reporter gene expression and cellular cytotoxicity was determined using luciferase and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, respectively. Acute toxicity in vivo was assessed by histological evaluation of harvested tissues. RESULTS Consistent activation of the MK and cox-2 promoters was noted in all of the ovarian cancer cell lines in addition to primary ovarian cancer cells. In contrast, reduced reporter activity was reported in mesothelial cells transduced with adenoviruses containing the test promoters, which was especially apparent for the cox-2 promoter. Additionally, the cox-2 promoter exhibited significantly lower reporter gene levels in liver and peritoneum than the control promoter in in vivo experiments. Tumor-cell killing induced by Adcox-2 MTK was comparable to that observed with AdCMVTK. However, a clear differential toxicity pattern was observed in favor of animals treated with Adcox-2 MTK when compared with controls. CONCLUSIONS These data clearly demonstrate that the transcriptional control afforded by the cox-2 promoter is tumor-specific and is able to mitigate associated toxicity in normal tissue while maintaining therapeutic efficacy in the context of an ovarian cancer molecular chemotherapeutic approach.