Midkine promoter-driven suicide gene expression and mediated adenovirus replication produced cytotoxic effects to immortalised and tumour cells
Midkine promoter-driven suicide gene expression and mediated adenovirus replication produced cytotoxic effects to immortalised and tumour cells
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DOI：
10.1016/j.ejca.2004.04.014
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发表时间：
2004-07-01
影响因子：
8.4
通讯作者：
Tagawa, A
中科院分区：
文献类型：
--
作者：
Yu, L;Hamada, K;Tagawa, A
我们研究了中期因子（MK）基因的调节区在癌症基因治疗中的可能应用，MK基因在许多人类肿瘤中经常上调，但在正常细胞中不上调。我们研究了MK基因组片段在成对细胞系、永生化细胞及其亲本正常成纤维细胞中的转录活性，发现MK片段优先在永生化细胞中激活融合的报告基因或自杀基因。将MK基因插入E1A基因上游的重组腺病毒（AdMK）在永生化细胞中优先复制，并对永生化细胞产生细胞毒性。与正常人成纤维细胞相比，人肝癌细胞对AdMK的敏感性显著增高，AdMK在感染细胞中的复制率低于野生型Ad。感染AdMK的肝细胞癌细胞在免疫功能低下的小鼠中没有形成肿瘤，并且将AdMK瘤内注射到小鼠中发展的肝细胞癌中延缓了随后的肿瘤生长。注射AdMK的病例中检测到E1A的表达和肿瘤坏死，而对照注射Ad的病例中未检测到E1A表达和肿瘤坏死。MK启动子驱动的自杀基因治疗和介导的复制型Ad可以对永生化细胞和肿瘤细胞产生细胞毒性作用，对正常细胞的损伤最小。(C)2004爱思唯尔有限公司保留所有权利。
We examined possible application of a regulatory region of midkine (MK) gene, which is frequently upregulated in a number of human tumours but not in normal cells, to cancer gene therapy. We examined transcriptional activity of the MK genomic fragments in paired cell lines, immortalized cells and their parental normal fibroblasts, and found that the MK fragments activated a fused reporter or a suicide gene preferentially in the immortalized cells. Recombinant adenoviruses (Ad), in which the MK fragment was inserted upstream to the E1A gene (AdMK), replicated preferentially in the immortalized cells and were cytotoxie to them. Human hepatocellular carcinoma cells were significantly susceptible to AdMK compared with human normal fibroblasts in vitro and the replication of AdMK was less than that of wild-type Ad in the infected fibroblasts. Hepatocellular carcinoma cells infected with AdMK did not form tumours in immunocompromised mice and intratumoural injection of AdMK into the hepatocellular carcinoma developed in mice retarded the subsequent tumour growth. Expression of E1A and necrosis of tumours were detected in AdMK-injected but not control Ad-injected cases. The MK promoter-driven suicide gene therapy and -mediated replicative Ad can thereby produce cytotoxic effects to immortalized and tumour cells with minimal damage to normal cells. (C) 2004 Elsevier Ltd. All rights reserved.