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Construction of FCU1 recombinant adenovirus vector and its killing effect on colon cancer cells
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- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 23
- DOI:
- 10.3969/j.issn.1009-3079.2008.23.007
- Key Word:
- 结肠癌;融合自杀基因;基因治疗
Abstract： 目的:构建含融合自杀基因FCU1的重组腺病毒载体,转染293细胞构建重组腺病毒,并感染结肠癌细胞,同时给以前体药物5-FC,观察其对结肠癌细胞的杀伤作用.方法:将FCU1基因采用EcoR Ⅰ、Sal Ⅰ双酶切克隆至穿梭质粒pDC316,构建质粒pDC316-FCU1,并酶切和测序鉴定.pDC316-FCU1与骨架质粒pBHG经脂质体共转染HEK 293细胞,在HEK 293细胞中同源重组并包装重组腺病毒Ad5-FCU1.将Ad5-FCU1感染结肠癌细胞HCT116,同时给以前体药物5-FC,MTT法检测5-FC对HCT116杀伤作用.结果:重组质粒PDC316-FCU1构建成功.pDC316-FCU1与骨架质粒pBHG共转染HEK293细胞后,出现明显的毒斑,纯化后病毒滴度为2×1012 PFU/L.重组病毒Ad5-FCU1感染HCT116细胞后,给以无毒前体药物5-FC显示极强的杀伤作用,当5-FC达100 μmol/L时,存活的HCT116/FCU1细胞极少.而对照组存活率接近90%.结论:成功构建了含融合自杀基因FCU1的Ad5-FCU1重组腺病毒,FCU1/5-FC自杀基因系统对结肠癌细胞具有显著的体外杀伤作用.
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