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，我们使用了基于二维凝胶电泳的限制性内切标记基因组扫描（RLGS）。这种方法有很多好处，其中之一是可以一次在一张x射线胶片上检测到数千个基因片段。该分析适用于人类结直肠癌和肝细胞癌。研究人员分析了这些癌症组织中特定的基因组变化，并确定了该基因片段的序列。此外，我们已经澄清了这些基因组改变与临床特征之间的相关性。1 ． 在一个结直肠癌病例中，二维凝胶电泳后发现DNA斑点平均有12个变化。提取斑点DNA片段，用ABI PRISM基因分析仪测定其核苷酸序列。在Gene Bank数据库中，该序列与人脂肪酸合成酶高度同源（65%），序列长度为410 bps。利用DNA点探针进行northern blot杂交，在仅有的癌组织中也观察到RNA的表达。在肝细胞癌病例中，DNA斑点平均有16个变化。RLGS变化点<大于等于>6个的患者的生存率明显低于RLGS变化点<小于等于>5个的患者。提取癌组织DNA改变的4个位点并进行测序。其中一个（1088 bps）位于8q21染色体，与人类串联重复序列一致。另一个（1415 bps）是着丝性NotI群集。肝细胞癌患者的无病生存率与肿瘤强度密切相关

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