探讨用表面增强基质辅助激光解析离子化一飞行时间质谱(SELDI-TOF-MS)筛选视网膜母细胞瘤(Retinoblastoma,Rb)血清标志蛋白,为Rb筛查、早期诊断及鉴别诊断寻找新的方法。 方法:采用IMAC30和CM102种蛋白芯片对18例Rb患者和17名年龄匹配的正常小儿血清进行了蛋白质谱检测。统计分析采用Ciphergenproteinchip3.0.2软件,2组间蛋白峰强度的比较采用t检验,率的比较采用Fisher精确概率法。 结果:IMAC30蛋白芯片结果显示:与正常幼儿相比,视网膜母细胞瘤患者血清中有26个差异表达的蛋白峰,其中高表达有21个,质/荷比(m/z)分别为:7746、7014、11713、3049、7084、7299、5888、2544、12575、5489、9658、9575、9929、10161、8955、1886、10617、6209、2411、7374和6614;低表达有5个,分别为:8382、7923、7972、8590和66576,统计学检验P值均小于0,001。其中以7014m/z峰进行两组统计学分析时,敏感性和特异性分别为94.4%和82.4%。与正常小儿相比,经CMIO蛋白芯片分析显示:视网膜母细胞瘤患者血清中有4个差异表达的蛋白峰,其中高表达有3个,分别为:质/荷比(m/z)分别为5888、6097和7798;低表达蛋白峰为8590m/z。统计学检验P值均小于0.叭。当选择7798m/z差异峰进行统计学分析时,敏感性和特异性分别是83.3%和70.6%。 结论:视网膜母细胞瘤患者血清中存在较多标志蛋白,SELDI-TOF-MS蛋白质芯片技术有望成为Rb筛查、早期诊断及鉴别诊断的新的有效工具。
Purpose: To find new serum biomarkers in the serum of retinoblastoma (Rb) patient with surface enhanced laser desorption/ ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and protein chip technique. Methods: SELDI-TOF-MS and IMAC30 and CM10 protein chips were used to analyze the protein profiles from serum of 18 patients with Rb and 17 age-matched controls. The protein profiling was analyzed statistically by Ciphergen protein chip software 3.0.2. The t-test was applied to compare the protein peak intensity. Fisher ratio-test was used to compare the predominance of differential protein peaks appeared in patients. Results: With IMAC30 protein chips, there were 26 proteins appearing differently in serum of patients with Rb compared to normal children. Among them, 21 proteins, i.e. 7 746, 70 146, 117 136, 30 496, 70 846, 72 996, 5 889, 2 544, 12 575, 5 489, 9658, 9575, 9929, 10 161, 8955, 1 886, 10617, 6209, 2411, 7374, 6614m/z were up-regulated and 5 proteins, i.e. 8 383, 7 923, 7 972, 8 590, 66 576 m/z, down-regulated(P〈 0.001 ). Using the 7014 m/z protein peak, we could differentiate the patients with Rb from the healthy children with a sensitivity of 94.4% and a specificity of 82.4%. By CM10 protein chips, 4 proteins, including 3 up-regulated proteins(5 890, 6 097, 7 798 m/z) and one down-regulated protein (8 593 m/z), were detected in Rb patients (P〈 0.01 ). The sensitivity and specificity were 83.3% and 70.6% respectively for 7 798 m/z protein. Conclusion : There are a few candidates of Rb biomarkers in the serum of Rb patient. SELDI Protein Chip technology could be a potential method in the clinical screening test of Rb.