in vitro manipulation of genes relevant to oncogenesis

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We have established the colon cancer cell lines in which the mutated K-ras gene was replaced by the normal K-ras gene, using homologous recombination. The comparative analyzes between these cell lines and the parental cell lines made it feasible to directly assess the role of the mutated K-ras gene in oncogenesis. As compared with the parental cell lines, the cell lines which lack the expression of the mutated K-ras gene showed the remarkable induction of C-fos and C-jun expression by the serum stimulation and the suppression of vascular endothelial growth factor (VEGF) production. These observations indicate that the mutated K-ras gene is involved in the suppression of the immediate early genes expression and the production of VEGF.Our strategy could be applied to clarify the role of the mutation in other oncogenes and tumor suppressor genes in oncogenesis.