Analysis of the protein markers and genetic markers to establish an early diagnostic systems for the oral cancer

1.口腔鳞状细胞癌组织中生物标志物的蛋白质组学分析：口腔鳞状细胞癌组织作为手术冰冻标本采集于Tsurumi大学附属医院8例患者的口腔鳞状细胞癌组织中，采用激光捕获显微切割方法获得同一个体的肿瘤和正常上皮组织。提取的蛋白质用SELDI TOF-MS系统进行差异蛋白质组分析，检测到明显增加或减少的峰型。这一结果表明，这些峰可以作为OSCC.2的候选生物标志物。口腔鳞癌危险诊断标记物(遗传标记物)的寻找：口腔鳞癌是一种遗传异常，遗传因素对其是否为癌构成有很大控制作用。因此，个人癌症风险诊断是实现个人癌症预防的必要手段。我们通过全基因组SNPs筛查的病例对照研究寻找了风险标记基因。收集和纯化DNA，用微球阵列系统检测SNPs。经卡方检验，2、6、7、8、10、11、12、16、19号染色体上的9个SNPs分别为0.0001、15和327个。接下来，我们利用EM算法进行了单倍型分型，结果表明1353个SNP参与了单倍型-块的形成，病例和对照之间的卡方检验结果表明，24个块的p值小于0.00001，但它们分散在染色体上；从1和2的结果来看，蛋白质和遗传标记有多个候选，它们的组合被认为是有意义的诊断标记。

1. Proteomic analysis for identification of the biomarkers in oral squamous cell carcinoma (OSCC) tissues:The tumor tissues of OSCC were collected as operation frozen-specimens from eight patients in the hospital attached to Tsurumi University, and the tumor and normal epithelial tissue of the same individuals were gained by the Laser Captured Microdissection method. The extracted proteins were analyzed by SELDI TOF-MS systems as differential proteome analysis, and the significantly increased or decreased peak patterns were detected. This result suggested these peaks should be the candidates for the biomarkers of OSCC.2. A search of risk diagnosis markers (genetic markers) of OSCC:The cancer is genetic abnormality, and it is thought that a hereditary factor controls it greatly whether it is cancer constitution. Therefore, an individual cancer risk diagnosis is necessary to realize the personal cancer prevention. We have searched the risk-marker genes by a case-control study of genome wide screening of the SNPs. The DNA was collected and purified, and the SNPs were detected by a micro-beads array systems. When the Chi-squared test was carried out on the difference of the distribution rate of 5984 SNPs between the 117 cases and 153 controls, 9SNPs of chromosome number 2,6,7,8,10,11,12,16,19 were shown as p<0.0001, 15SNPs were p<0.01, and 327SNPs were p<0.05. Next, we carried out the haplotyping using the EM algorism, it was shown that the 1353 SNPs participated in the haplotype-block formation, and the result of Chi-squared test between cases and controls showed the p-values of 24 blocks were less than 0.00001, but those were scattered on chromosomes.From the results of 1 and 2, it was shown that there are plural candidates of protein and genetic markers, and it was thought the combination of them might make significant as diagnostic markers.