IN-VIVO ADENOVIRUS-MEDIATED GENE-TRANSFER OF THE ESCHERICHIA-COLI CYTOSINE DEAMINASE GENE TO HUMAN COLON CARCINOMA-DERIVED TUMORS INDUCES CHEMOSENSITIVITY TO 5-FLUOROCYTOSINE
IN-VIVO ADENOVIRUS-MEDIATED GENE-TRANSFER OF THE ESCHERICHIA-COLI CYTOSINE DEAMINASE GENE TO HUMAN COLON CARCINOMA-DERIVED TUMORS INDUCES CHEMOSENSITIVITY TO 5-FLUOROCYTOSINE
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DOI：
10.1089/hum.1995.6.8-1055
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发表时间：
1995-08-01
影响因子：
4.2
通讯作者：
CRYSTAL, RG
中科院分区：
文献类型：
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作者：
HIRSCHOWITZ, EA;OHWADA, A;CRYSTAL, RG
为了评估大肠杆菌胞嘧啶脱氨酶基因的体内转移是否会赋予实体肿瘤对前药5-氟胞嘧啶（5FC）的敏感性，我们构建了一种由巨细胞病毒（CMV）启动子驱动的携带胞嘧啶脱氨酶基因的腺病毒载体（AdCMV.CD），在体外和体内感染HT29结肠癌细胞，并评估细胞随时间的生长情况。AdCMV。CD在体外HT29细胞中产生功能性胞嘧啶脱氨酶蛋白，感染细胞的裂解物能够将[H-3]5FC转化为其活性代谢物5-氟尿嘧啶（5FU）。AdCMV。在5FC存在的情况下，CD载体能有效抑制HT29细胞的体外生长，并呈剂量依赖性。AdCMV感染。在细胞混合研究中，当只有10%的细胞表达胞嘧啶脱氨酶基因时，CD与5FC联合使用时会产生旁观者效应。此外，这种旁观者效应并不依赖于细胞间的接触，正如从AdCMV上清液中抑制HT29细胞中[H-3]胸苷结合所证明的那样。用5FC处理cd感染的细胞转移到未感染的细胞。与这些体外观察结果一致，当AdCMV。将CD直接注射到接受5FC治疗的裸鼠HT29皮下肿瘤中，与对照组相比，第15天肿瘤大小缩小了4倍，第28天缩小了5倍。这些观察结果表明，腺病毒介导的大肠杆菌胞嘧啶脱氨酶基因的基因转移和伴随的5FC的施用可能是一种局部控制对5FU敏感的肿瘤细胞生长的策略。
To evaluate the concept that in vivo transfer of the Escherichia coli cytosine deaminase gene will confer sensitivity of a solid tumor to the prodrug 5-fluorocytosine (5FC), we constructed an adenovirus vector (AdCMV.CD) carrying the cytosine deaminase gene driven by the cytomegalovirus (CMV) promoter, infected HT29 colon carcinoma cells in vitro and in vivo, and evaluated cell growth over time. AdCMV.CD produced a functional cytosine deaminase protein in HT29 cells in vitro as evidenced by the ability of lysates from the infected cells to convert [H-3]5FC to its active metabolite 5-fluorouracil (5FU). The AdCMV.CD vector effectively suppressed HT29 cell growth in vitro in the presence of 5FC in a dose-dependent manner. Infection with AdCMV.CD, when as few as 10% of cells expressed the cytosine deaminase gene, was associated with a bystander effect when combined with 5FC in cell mixing studies. Further, this bystander effect was not dependent on cell-to-cell contact as demonstrated by suppression of [H-3]thymidine incorporation in HT29 cells when supernatant from AdCMV.CD-infected cells treated with 5FC was transferred to uninfected cells. Consistent with these in vitro observations, when AdCMV.CD was directly injected into established subcutaneous HT29 tumors in nude mice receiving 5FC, there was a four-fold reduction in tumor size at day 15 compared to controls, and a five-fold reduction at day 28. These observations suggest that adenovirus-mediated gene transfer of the E. coli cytosine deaminase gene and concomitant administration of 5FC may have potential as a strategy for local control of the growth of tumor cells susceptible to 5FU.