Hargest R, Williamson R
Department of Biochemistry and Molecular Genetics, St Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London.
Gut. 1995 Dec;37(6):826-9. doi: 10.1136/gut.37.6.826.
Mutations in the adenomatous polyposis coli (APC) gene cause the hereditary cancer syndrome familial adenomatous polyposis and are implicated in the early stages of sporadic colorectal carcinogenesis. APC is therefore a promising candidate for use in prophylactic gene therapy of intestinal tissues at high risk of becoming malignant. The aim of the study was to discover if functional full length APC gene can be introduced into somatic gut epithelial cells and to define the optimum conditions for such transfer. Copies of the normal APC gene were introduced into SW480 cells, a colonic epithelial cell line with an APC gene mutation, using plasmid DNA combined with liposomes. Reverse transcriptase polymerase chain reaction and restriction enzyme digestion allowed the endogenous gene to be distinguished from the transgene. It was shown that the normal APC gene is expressed at high levels for 72 hours after transfection and disappears within one week. This study shows that short-term expression of normal APC gene can be achieved after transfection with liposome-DNA complexes at sufficiently high levels to permit assessment of biological effects.
腺瘤性结肠息肉病（APC）基因的突变会导致遗传性癌症综合征——家族性腺瘤性息肉病，并且与散发性结直肠癌发生的早期阶段有关。因此，APC是用于对有恶变高风险的肠道组织进行预防性基因治疗的一个很有前景的候选基因。本研究的目的是探究功能性全长APC基因能否被导入体肠道上皮细胞，并确定这种转移的最佳条件。使用与脂质体结合的质粒DNA，将正常APC基因的拷贝导入SW480细胞（一种具有APC基因突变的结肠上皮细胞系）。逆转录聚合酶链反应和限制性酶切可将内源基因与转基因区分开来。结果表明，正常APC基因在转染后72小时高水平表达，并在一周内消失。这项研究表明，用脂质体-DNA复合物转染后，正常APC基因能够实现短期表达，其表达水平足以评估生物学效应。
