In vivo human carboxylesterase cDNA gene transfer to activate the prodrug CPT-11 for local treatment of solid tumors
In vivo human carboxylesterase cDNA gene transfer to activate the prodrug CPT-11 for local treatment of solid tumors
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DOI：
10.1172/jci119888
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发表时间：
1998-04-15
影响因子：
15.9
通讯作者：
Crystal, RG
中科院分区：
文献类型：
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作者：
Kojima, A;Hackett, NR;Crystal, RG
为了评估体内转移人羧酸酯酶基因将赋予实体瘤对前药CPT-11（伊立替康）的敏感性的概念，我们构建了携带由巨细胞病毒（CMV）启动子驱动的人羧酸酯酶基因的腺病毒载体（AdCMV.CE），在体外和体内感染A549人肺腺癌细胞，并评估细胞生长Dyer时间。AdCMV.CE在体外和体内在A549细胞中产生功能性羧酸酯酶蛋白，如通过来自感染细胞的裂解物将CPT-11转化为其活性代谢物SN-38的能力所证明的。在CPT-11存在下，AdCMV.CE载体在体外有效抑制A549细胞生长。细胞混合研究表明，当只有10%的细胞表达人羧酸酯酶基因时，在CPT-11存在下存在旁观者生长抑制。与这些体外观察结果一致，当将AdCMV.CE直接注射到接受CPT-11的裸鼠中建立的皮下A549肿瘤中时，与对照相比，在第27天肿瘤大小减少35%，在第34天减少41%（P < 0.01，均与对照相比）。用细胞系H157和HeLa进行了类似的观察，这些观察结果表明，人羧酸酯酶基因的局部基因转移和伴随的CP=T-11的局部施用可能具有作为控制实体瘤生长的策略的潜力。
To evaluate the concept that in vivo transfer of the human carboxylesterase gene will confer sensitivity of a solid tumor to the prodrug CPT-11 (irinotecan), we constructed an adenovirus vector (AdCMV.CE) carrying the human carboxylesterase gene driven by the cytomegalovirus (CMV) promoter, infected A549 human lung adenocarcinoma cells in vitro and in vivo, and evaluated cell growth Dyer time. AdCMV.CE produced a functional carboxylesterase protein in A549 cells in vitro and in vivo as evidenced by ability of lysates from the infected cells to convert CPT-11 to its active metabolite SN-38. The AdCMV.CE vector effectively suppressed A549 cell growth in vitro in the presence of CPT-11. Cell mixing studies demonstrated that when as few as 10% of cells expressed the human carboxylesterase gene, there was bystander growth suppression in the presence of CPT-11. Consistent with these in vitro observations, when AdCMV.CE was directly injected into established subcutaneous A549 tumors in nude mice receiving CPT-11, there was 35% reduction in tumor size at day 27 compared to controls, and a 41% reduction at day 34 (P < 0.01, both comparisons to controls). Similar observations were made with the cell line H157 and HeLa, These observations suggest that local gene transfer of the human carboxylesterase gene and concomitant local administration of CP=T-11 may have potential as a strategy for control of the growth of solid tumors.