论文部分内容阅读
探讨用蛋白质芯片技术筛选肺癌病人血清中的特异性标志蛋白。采用SELDI(surfaced enhanced laser desorption/ionization)蛋白芯片技术检测了30例肺癌(15例原发癌,9例转移癌,6例化疗后)病人血清和12例正常人血清中的蛋白质谱。选用阴离子交换柱处理血清样品,用5个不同pH的洗脱液和有机溶剂洗脱柱床,通过改变洗脱液的pH值得到6个具有不同pI的蛋白质组分,每个组分用IMAC-Cu和WCX2两种芯片检测。采用美国Ciphergen生物公司研制的蛋白质芯片阅读机读取数据,获得的蛋白质谱采用Ciphergen公司的Biomark Wizard软件分析。结果显示,肺癌病人与正常人的血清中有15个差异蛋白(标志分子)表达。与正常人血清相比,6个蛋白质在肺癌病人血清中高表达,9个蛋白质在肺癌病人血清中低表达,采用单一标志分子检测时,敏感性为44.82％～93.1％,特异性为85％～94.4％。其中5个标志分子在两种芯片上被检测出来。实验证明与肺癌相关的特异性标志分子可从病人血清中检测出来,而且蛋白质芯片技术对于发现和筛选血清中的肺癌标志蛋白是一种有效、快速的工具。
To explore the use of protein chip technology screening of lung cancer patients serum specific marker protein. The protein profiles of serum from 30 patients with lung cancer (15 primary carcinomas, 9 metastatic carcinomas, 6 with chemotherapy) and 12 normal controls were examined using SELDI (surfaced enhanced laser desorption / ionization) protein chip technology. Separation of serum samples by anion exchange column, elution with five different pH eluents and organic solvent, and changing of the pH of the eluate resulted in six protein fractions with different pI, each with IMAC -Cu and WCX2 two chip detection. Data were read using a protein chip reader developed by Ciphergen Bio Inc., USA. Protein profiles obtained were analyzed using Biomark Wizard software from Ciphergen. The results showed that there were 15 differentially expressed proteins (marker molecules) in serum of lung cancer patients and normal people. Compared with normal human serum, 6 proteins were highly expressed in sera of patients with lung cancer, and 9 proteins were low in sera of patients with lung cancer. Sensitivity was 44.82% ~ 93.1% and specificity was 85% ~ 94.4%. Five of the marker molecules were detected on both chips. Experiments have shown that specific markers associated with lung cancer can be detected from the patient’s serum, and protein chip technology is an effective and rapid tool for the detection and screening of lung cancer marker proteins in serum.