摘要
目的:利用质谱技术和生物信息学方法从前列腺癌患者血清中筛选并鉴定血清淀粉样蛋白A(SAA)。方法:应用SELDI技术检测前列腺癌差异蛋白的表达变化,筛选11.1～11.9kD的一簇蛋白峰。采用HPLC技术分离该蛋白,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析得到相应的肽质指纹图谱,然后搜索数据库鉴定部分差异蛋白质点。结果:分析显示11.1～11.9kD的一簇蛋白峰在前列腺癌患者血清中明显高于对照组,该蛋白峰的表达水平与前列腺癌患者的临床分期密切相关,随着病情的加重而逐渐升高。HPLC结合MALDI技术鉴定出芯片上该簇蛋白峰为SAA,并得到ELISA方法的验证。结论:质谱技术可作为检测前列腺癌生物标记的方法,具有易操作、敏感性高的特点。本研究发现的这组蛋白标记物可能对前列腺癌的诊断及疗效观察具有潜在的应用价值。
Objective： To screen and identify serum amyloid A （SAA） in patients with prostate cancer with mass spectrum technique. Methods： SELDI technology was used to detect the changes in protein expression. SAA was screened and separated and then identified by peptide mass fingerprint （PMF） based on matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and database searching. Results： The analysis showed that a mass cluster in the ranges of 11.1-11.9KD in M/Z value in the serum of prostate cancer patients was much higher than that in the control group. Meanwhile, this protein peak was closely correlated with clinical stages of prostate cancer. The level of the protein peak was increased as the illness got serious. Through MALDI technology combined with HPLC, the mass cluster in the range of 11.1-11.9KD in M/Z value on the chip was identified as SAA. And it was also verified through ELISA method. Conclusion： Mass spectrum technology is an effective method to detect the biological markers in prostate cancer patients. This method is convenient, highly sensitive and with good reproducibility. The SAA can be used as a marker in the diagnosis of prostate cancer. These indices are also meaningful in screening and identifying signal proteins from the serum of prostate cancer patients.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2009年第21期1227-1231,共5页
Chinese Journal of Clinical Oncology
基金
浙江省医药卫生科研基金资助(编号:2006A123)
