Identification of new oncogenes related to human cancer by human genomic DNA analysis based on two-dimensio electrophoresi

为了评估与人类癌症相关的基因组DNA的改变，我们使用基于二维凝胶电泳的限制性标志基因组扫描（RLGS）。这种方法有很多好处，其中之一是可以在一张X光片上同时检测出数千个基因片段。该分析应用于人结直肠癌和肝细胞癌。分析了这些癌组织中特异性的基因组改变，并鉴定了基因片段的序列。此外，我们还阐明了这些基因组改变与临床特征之间的相关性。1例大肠癌患者经双向电泳后，平均有12个DNA斑点改变。提取其中一个DNA片段，用ABI PRISM GENETIC ANALYSER进行核苷酸序列测定，发现该片段与基因库中人脂肪酸合成酶的同源性为65%，为410 bp。用DNA斑点探针进行北方杂交，在癌组织中也观察到RNA的表达.平均每例肝癌有16个DNA斑点改变。有16个RLGS点改变的患者的生存率<greater than or equal>显著低于RLGS点改变较少（15<less than or equal>个点）的患者。提取癌组织中的4个DNA突变点并测序。一个（1088 bp）位于染色体8q21，与人类串联重复序列相对应。另一个（1415 bp）为NotI簇。肝细胞癌患者的无病生存率与这些指标的强度密切相关，

To evaluate alterations of genomic DNAs related to human cancer, we used restriction landmark genomic scanning (RLGS) based on two-dimensional gel electrophoresis. This method has many benefits, one of which can detect thousands of gene fragments on one X-ray film at one time. This analysis was applied for human colorectal cancer and hepatocellular carcinoma. Genomic alterations specific in those cancer tissues were analyzed and the sequences of the gene fragment were identified. Furthermore, we have clarified the correlation between those genomic alterations and clinical characteristics.1 . Average twelve changes in DNA spots after two-dimensional gel electrophoresis were seen in a case with colorectal cancer. one of the DNA fragments in the spot was extracted and the nucleotide sequence was determined by ABI PRISM GENETIC ANALYSER.The sequence (410 bps) was found to be highly homologous to human fatty acid syhthase (65 %) in Gene Bank database. The expression of the RNA was also observed in the only cancer tissues by northern blot hybridization using the probe from the DNA spot.2. Average sixteen changes in DNA spots were seen in a case with hepatocellular carcinoma. The survival rates for patients with <greater than or equal> 16 changed RLGS spots were significantly lower than those for patients with fewer changed RLGS spots ( <less than or equal> 15 spots). Four spots of the DNA alterations of cancer tissues were extracted and sequenced. One (1088 bps) was located at chromosome 8q21 and corresponded to human tandem repeat sequence. Another one (1415 bps) was centromeric NotI cluster. The disease-free survival rates for patients with hepatocellular carcinoma were closely associated with the intensity of those