The use of the L-plastin promoter for adenoviral-mediated, tumor-specific gene expression in ovarian and bladder cancer cell lines.
The use of the L-plastin promoter for adenoviral-mediated, tumor-specific gene expression in ovarian and bladder cancer cell lines.
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发表时间：
2001-06
期刊：
影响因子：
11.2
通讯作者：
X. Peng;J. Won;T. Rutherford;T. Fujii;D. Zelterman;G. Pizzorno;E. Sapi;J. Leavitt;B. Kacinski;R. Crystal;P. Schwartz;A. Deisseroth
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文献类型：
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作者：
X. Peng;J. Won;T. Rutherford;T. Fujii;D. Zelterman;G. Pizzorno;E. Sapi;J. Leavitt;B. Kacinski;R. Crystal;P. Schwartz;A. Deisseroth
将2.4-kb截短的L-质体启动子插入到LacZ基因的5'端（Ad-Lp-LacZ）或胞嘧啶脱氨酶（CD）基因的5'端（Ad-Lp-CD）中的无复制能力的腺病毒载体骨架中。与LacZ和CD载体的生物活性和细胞毒性实验表明，L-质体启动子驱动的转录单位表达在更高的水平，从患者的卵巢癌细胞的外植体，并在建立卵巢癌或膀胱癌细胞系比他们在正常的腹膜间皮瘤细胞从手术标本，在器官培养的正常卵巢细胞，或在建立CCD最小偏差成纤维细胞系。对照实验表明，这种差异不是由于缺乏感染性的正常腹膜细胞，正常卵巢细胞，或最小偏差CCD成纤维细胞系，因为这些细胞表现出表达的LacZ报告基因时，暴露于携带的巨细胞病毒（CMV）驱动的LacZ基因（Ad-CMV-LacZ）的复制缺陷型腺病毒载体。暴露于Ad-Lp-CD腺病毒载体的Ovcar-5和Skov-3卵巢癌细胞系比暴露于相同载体后的CCD最小偏差成纤维细胞系对前药5-氟胞嘧啶（5 FC）更敏感，5 FC从5 FC前药转化为毒性化学物质5-氟尿嘧啶。采用小鼠卵巢癌移植瘤模型，观察Ad-Lp-CD载体/5 FC系统对卵巢癌细胞裸鼠移植瘤的抑制作用。最后，将Ad-Lp-CD载体注射到s.c.肿瘤结节产生的肿瘤结节大小的减小比将Ad-CMV-LacZ载体注射到肿瘤结节中产生的肿瘤结节大小的减小更大。Ad-Lp-CD载体与Ad-CMV-CD载体对肿瘤生长具有相同的抑制作用。这些结果表明，携带由L-plastin启动子控制的CD基因的腺病毒载体（Ad-Lp-CD）可能对卵巢癌的腹腔注射治疗具有潜在价值。
A 2.4-kb truncated L-plastin promoter was inserted either 5' to the LacZ gene (Ad-Lp-LacZ) or 5' to the cytosine deaminase (CD) gene (Ad-Lp-CD) in a replication-incompetent adenoviral vector backbone. Infectivity and cytotoxicity experiments with the LacZ and CD vectors suggested that the L-plastin promoter-driven transcriptional units were expressed at much higher levels in explants of ovarian cancer cells from patients and in established ovarian or bladder cancer cell lines than they were in normal peritoneal mesothelial cells from surgical specimens, in organ cultures of normal ovarian cells, or in the established CCD minimal deviation fibroblast cell line. Control experiments showed that this difference was not attributable to the lack of infectivity of the normal peritoneal cells, the normal ovarian cells, or the minimal deviation CCD fibroblast cell line, because these cells showed expression of the LacZ reporter gene when exposed to the replication-incompetent adenoviral vector carrying the cytomegalovirus (CMV)-driven LacZ gene (Ad-CMV-LacZ). The Ovcar-5 and Skov-3 ovarian cancer cell lines exposed to the Ad-Lp-CD adenoviral vector were much more sensitive to the prodrug 5-fluorocytosine (5FC), which is converted from the 5FC prodrug into the toxic chemical 5-fluorouracil, than was the CCD minimal deviation fibroblast cell line after exposure to the same vector. A mouse xenograft model was used to show that the Ad-Lp-CD vector/5FC system could prevent engraftment of ovarian cancer cells in nude mice. Finally, injection of the Ad-Lp-CD vector into s.c. tumor nodules generated a greater reduction of the size of the tumor nodules than did injection of the Ad-CMV-LacZ vectors into tumor nodules. The Ad-Lp-CD vectors were as suppressive to tumor growth as the Ad-CMV-CD vectors. These results suggest that an adenoviral vector carrying the CD gene controlled by the L-plastin promoter (Ad-Lp-CD) may be of potential value for the i.p. therapy of ovarian cancer.