Nagata Y, Abe M, Kobayashi K, Saiki S, Kotake T, Yoshikawa K, Ueda R, Nakayama E, Shiku H
Department of Oncology, Nagasaki University School of Medicine.
Jpn J Cancer Res. 1990 Jan;81(1):22-7. doi: 10.1111/j.1349-7006.1990.tb02502.x.
Point mutations of c-ras genes at codons 12, 13 and 61 were analyzed in 26 cases of bladder cancer and 16 cases of kidney cancer. DNA prepared from either frozen tissues or 10% formalin-fixed, paraffin-embedded tissues were amplified by means of polymerase chain reaction methods, and mutations were analyzed by dot blot hybridization assays with oligonucleotide probes. In three cases of bladder cancer c-ras mutations were found, at codons 13 and 61 of c-Ha-ras and at codon 61 of c-Ki-ras, while no mutation was found in kidney cancer. No mutation was found in normal bladder epithelial tissues from the same patients. Our findings, taken together, may indicate relative scarcity of c-ras mutations in these types of human cancer. The results of dot blot hybridization assays and DNA sequencing showed a G-to-C transition of the first nucleotide at codon 13 c-Ha-ras. This is the first time that such a point mutation has been detected in human cancer tissues.
对26例膀胱癌和16例肾癌患者分析了c-ras基因第12、13和61位密码子的点突变情况。采用聚合酶链反应方法对取自冷冻组织或10%福尔马林固定、石蜡包埋组织的DNA进行扩增,并用寡核苷酸探针通过斑点杂交试验分析突变情况。在3例膀胱癌中发现了c-ras突变,分别位于c-Ha-ras的第13和61位密码子以及c-Ki-ras的第61位密码子,而在肾癌中未发现突变。在同一患者的正常膀胱上皮组织中也未发现突变。综合我们的研究结果,可能表明在这些类型的人类癌症中c-ras突变相对较少。斑点杂交试验和DNA测序结果显示,c-Ha-ras第13位密码子的第一个核苷酸发生了G到C的转换。这是首次在人类癌组织中检测到这种点突变。