乳头状甲状腺癌患者血清中特异性标志物的检测与鉴定

目的 探讨并鉴定乳头状甲状腺癌患者血清中肿瘤相关蛋白作为特异性标志物的可能性.方法 应用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测35例乳头状甲状腺癌、40例甲状腺良性结节和34例健康对照者的血清标本,应用生物信息学方法筛选差异蛋白峰,经高效液相色谱(HPLC)分离出差异蛋白,酶解后进行液质联用串联质谱(LC-MS/MS)分析,用SEQUEST检索程序查询Bioworks数据库蛋白序列进行鉴定.结果 筛选出6个最显著的差异蛋白质,质荷比(M/Z)分别位于6651、6452、7653、7932、15 106和15 848.M/Z位于6651、6452处的蛋白质在乳头状甲状腺癌组表达低于甲状腺良性结节和健康对照组;M/Z位于7653、7932、15 106、15 848处的蛋白质在乳头状甲状腺癌组表达高于甲状腺良性结节和健康对照组,差异均有统计学意义(均P<0.01).联合6种潜在蛋白质标志物,区别乳头状甲状腺癌和非乳头状甲状腺癌的特异度为88.0%,敏感度为92.5%.bl/Z位于6651、6452、7653和15 106、7932和15 848处的蛋白标志物分别为载脂蛋白C-Ⅰ、载脂蛋白C一Ⅲ、α-珠蛋白、β-珠蛋白.结论 鉴定出的载脂蛋白C-Ⅰ、载脂蛋白C-Ⅲ、α-珠蛋白和β-珠蛋白在乳头状甲状腺癌的诊断中具有一定的价值和广泛的应用前景,值得进一步研究和探讨.

Detection and identification of specific serum biomarkers in papillary thyroid cancer

Objective To detect and identify the potential specific serum biomarkers for diagnosis of papillary thyroid cancer. Methods Samples of 35 patients with papillary thyroid carcinoma, 40 patients with benign thyroid nodule and 34 healthy individuals were analyzed using the SELDI-TOF ProteinChip System and bioinfomation technology to find the differential peaks which were separated by HPLC and then further analyzed by LC-MS/MS. The protein sequences were analyzed by SEQUEST software and searched in Bioworks database. Results The top six mass-to-charge ratio (M/Z) peaks with the smallest P value were 6651,6452, 7653, 7932, 15 106 and 15 848 Da, respectively. The 6651 and 6452 Da proteins were weakly expressed in papillary thyroid carcinoma but highly expressed in benign thyroid nodules and healthy individuals. The differences had statistical significance (P<0.01). The 7653, 7932, 15 106, 15 848 Da proteins were highly expressed in papillary thyroid carcinoma but weakly expressed in benign thyroid nodules and healthy individuals. The differences were statistically significant(P<0.01). Combination of these six proteins, using the method of leave-one-out to make crossing detection, the specificity of discriminating papillary thyroid carcinoma and non-cancer was 88.0%, and its sensitivity was 92.5%. The 6651 and 6452 Da proteins were identified as apolipoprotein C-Ⅰ and apolipoprotein C-Ⅲ, respectively. The 7653 and 15 106 Da proteins were identified as the same pretein-α-globin, and the 7932 and 15 848 Da proteins were identified as the same protein-β-globin. Conclusion The detection of differentially expressed apolipoprotein C-Ⅰ, apolipoprotein C-Ⅲ, α-globin, and β-globin may have utility for diagnosis of papillary thyroid carcinoma and are worthy of further investigation.