目的:探讨用表面增强基质辅助激光解析离子化-飞行时间质谱(SELDI-TOF-MS)筛选视网膜母细胞瘤(retinoblas-toma,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CM10两种蛋白芯片对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,两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〈0.01。其中以m/z7014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CM10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,质/荷比(m/z)分别为5888,6097和7798;低表达蛋白峰为m/z8590。统计学检验P〈0.01。当选择m/z7798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。
AIM: To find new biomarkers in the sera of retinoblastoma (Rb) patients with surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and protein chip technique. METHODS: SELDI-TOF-MS, IMAC30 and CM10 protein chips were used to analyze the protein profiles from sera 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's exact test was used to compare the predominance of differential protein peaks appeared in patients. RESULTS: With IMAC30 protein chips, there were 26 proteins which appeared different in sera of patients with Rb compared to normal children. Among them, 21 proteins, i. e. 7746, 7014, 11713, 3049, 7084, 7299, 5888, 2544, 12575, 5489, 9658, 9575, 9929, 10161, 8955, 1886, 10617, 6209, 2411,7374, 6614 m/z were up-regulated and 5 proteins, i. e. 8382, 7923, 7972, 8590, 66576 m/z, were down-regulated(P〈0. 01). Using the 7014 m/z protein peak for statistical analysis, 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 (5888, 6097, 7798 m/z) and 1 down-regulated protein (8590 m/z), were detected in Rb patients (P〈0.01 ). The sensitivity and specificity were 83. 3% and 70. 6% respectively when 7798 m/z protein peak was selected for statistical analysis. CONCLUSION: There are a few candidates as Rbbiomarkers in the sera of Rb patients. SELDI-TOF-MS protein chip technology could be a potential method in the clinical screening test of Rb.