Expression of the APC gene after transfection into a colonic cancer cell line.
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.
Hargest
R
R
Department of Biochemistry and Molecular Genetics, St Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London.
Williamson
R
R
D001483:Base Sequence; D015179:Colorectal Neoplasms / Q000235:genetics*; D015870:Gene Expression; D017491:Genes, APC; D015316:Genetic Therapy; D006801:Humans; D008081:Liposomes; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014162:Transfection; D014407:Tumor Cells, Cultured
D008081:Liposomes