骨肉瘤血清蛋白标志物的初步研究

 目的 利用 Link-test统计方法整合分析基因芯片和蛋白质质谱分析数据寻找骨肉瘤血清蛋白标志物。方法利用基因芯片检测从骨肉瘤细胞系(MG-63、Saos-2和 U-2 OS)中筛选骨肉瘤候选标志基因；运用蛋白质质谱分析技术分析 27个骨肉瘤患者血清标本及 47个健康人的血清标本, 从中筛选差异具有统计学意义的蛋白质质谱峰；采用 Link-test统计方法对筛选到的骨肉瘤候选标志基因和蛋白质质谱峰进行相关性分析, 得到交叉验证的骨肉瘤血清蛋白标志物；运用免疫印迹法比较一种候选血清蛋白标志物在骨肉瘤患者和健康人血清中的含量差异。结果利用基因芯片检测到 653个候选骨肉瘤标志基因, 蛋白质质谱分析筛选到 6个蛋白质质谱峰。经 Link-test统计分析, 共鉴定出 13个候选骨肉瘤血清蛋白标志物, 其中包括细胞色素 C(Cytochrome c1, CYC-1)。在 13个候选蛋白质标志物中, 12个在骨肉瘤中表达上调, 1个表达下调。免疫印迹实验发现 CYC-1在骨肉瘤患者血清中的含量明显高于健康人, 差异具有统计学意义。结论利用 Link-test统计方法整合分析基因芯片和蛋白质质谱分析数据是寻找骨肉瘤血清蛋白标志物的有效方法, 共发现 13个候选骨肉瘤血清蛋白标志物。

Primary study on candidate serum markers of osteosarcoma

Objective To discover potential serum biomarkers of osteosarcoma by an integrated analysis of datasets from gene chip and surface enhanced laser desorption-ionization time-off-light mass spectrometry (SELDI-TOF-MS).Methods Gene chip was used to screen potential biomarkers of osteosarcoma from osteosarcoma cell lines (MG-63,Saos-2 and U-2OS),and SELDI-TOF-MS was used to screen protein peaks with statistically significant difference from serum samples (27 osteosarcoma cases and 47 healthy cases).The association between these two types of markers was constructed by link-test to get cross-validated serum biomarkers of osteosarcoma.Immunoblot analysis was performed to detect the expression of one candidate osteosarcoma protein maker in serums from osteosarcoma patients and members of the control group.Results Six hundred and fifty three candidate osteosarcoma gene markers were found by gene chip and 6 protein peaks were found by SELDI-TOF-MS.Thirteen differentially expressed serum markers of osteosarcoma including CYC-1 were screened by link-test analysis (12 of them were up-regulated in serum samples of the osteosarcoma patients while 1 was down-regulated ).By immunoblot,we found that CYC-1 protein was significantly up-regulated in serums from osteosarcoma patients compared to the control group.Conclusion An integrated analysis of datasets from gene chip and SELDI-TOF-MS could accelerate the discovery,screening and identification of tumor biomarkers.A total of 13 potential serum protein markers of osteosarcoma were discovered in this study.