Yuasa Y, Oto M, Sato C, Miyaki M, Iwama T, Tonomura A, Namba M
Jpn J Cancer Res. 1986 Sep;77(9):901-7.
The DNA of a colon carcinoma-derived cell line (KMS-4) and that of skin fibroblasts from a familial polyposis coli patient were transfected into NIH3T3 cells in order to detect oncogenes associated with the disease. No transformation was observed with the normal skin fibroblast DNA, while the KMS-4 cell DNA was able to transform NIH3T3 cells. Through hybridization with known oncogene probes, the KMS-4 transforming gene was found to be a human activated c-K-ras 2 oncogene. Sequence analysis of the molecularly cloned KMS-4 c-K-ras 2 oncogene showed a single nucleotide transition from G to T at the 12th codon. This results in substitution of cysteine for glycine at this position. On using labeled synthetic oligonucleotides to detect the mutation in codon 12, we found the G to T transition in colon carcinoma cells. This suggests that activation of the c-K-ras 2 oncogene could be associated with colon carcinoma induction.
为了检测与该疾病相关的致癌基因，将结肠癌细胞系（KMS - 4）的DNA和来自家族性结肠息肉病患者的皮肤成纤维细胞的DNA转染到NIH3T3细胞中。正常皮肤成纤维细胞DNA未观察到转化现象，而KMS - 4细胞DNA能够转化NIH3T3细胞。通过与已知致癌基因探针杂交，发现KMS - 4转化基因是人类活化的c - K - ras 2致癌基因。对分子克隆的KMS - 4 c - K - ras 2致癌基因进行序列分析，发现在第12密码子处有一个从G到T的单核苷酸转换。这导致该位置的甘氨酸被半胱氨酸取代。在用标记的合成寡核苷酸检测第12密码子的突变时，我们在结肠癌细胞中发现了从G到T的转换。这表明c - K - ras 2致癌基因的激活可能与结肠癌的诱发有关。
