基因组学与应用生物学

为探究胃癌患者血清中可能存在的特异性蛋白标记物,本研究选取贵州省骨科医院的80名胃癌组患者和80名对照组人员(40名健康志愿者,40名慢性萎缩性胃炎患者)作为研究对象,采用表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)法筛选血清蛋白标记物,然后用HPLC法筛选和分离不同表达的蛋白峰,经酶解后进行液相色谱质谱/质谱联用(LC-MS/MS)分析,再用Bio Work 3.2进行数据分析,采用免疫印迹检测胃癌组和对照组血清蛋白标记物的表达。结果显示,在胃癌组患者血清中发现三个表达不同的蛋白峰,分别为m/z 5 905.6、m/z 6 638.5和m/z 8 710.3。相比对照组,胃癌组的m/z 5905.6峰表达信号增强(胃癌组为(7.95±3.32),对照组为(0.77±0.21)),而m/z 6 638.5蜂(胃癌组为(5.78±2.55),对照组为(16.45±4.32))和m/z 8 710.3峰(胃癌组(0.94±0.19),对照组为(2.23±0.87))的表达信号减弱。利用m/z 5 905.6、m/z 6 638.5和m/z 8 710.3三个峰的表达信号差别,可确定胃癌患者血清中的特异性的蛋白标记物,并利用这3种特异性的蛋白标记物对胃癌进行检测,其中胃癌组和对照组的检出率分别为93.3%和95%。蛋白鉴定结果显示,m/z 5905.6、m/z 6 638.5和m/z 8 710.3三个蜂分别是纤维蛋白原α链(fibrinogen alpha chain,FGA),载脂蛋白A-Ⅱ(apolipoprotein A-Ⅱ,Apo A-Ⅱ)和载脂蛋白C-Ⅰ(apolipoprotein C-Ⅰ,Apo C-Ⅰ);Western blotting结果显示胃癌患者血清FGA表达显著升高(p<0.05),Apo A-Ⅰ和Apo C-Ⅰ的表达显著降低(p<0.05)。研究数据表明,FGA、Apo A-Ⅱ和Apo C-Ⅰ这3种血清蛋白标记物在胃癌中的表达程度与胃癌进展程度有关,临床应用上具有潜在的诊断价值。

In order to explore the possible specific protein markers in the serum of gastric cancer patients, 80 patients with gastric cancer in Guizhou Osteological Hospital and a control group with 80 people(40 healthy volunteers and 40 patients with chronic atrophic gastritis) were selected as the research subjects, and the surface enhanced laser desorption ionization time of flight mass spectrometry(SELDI-TOF-MS) was used. The serum protein markers were screened by the method, then the protein peaks of different expressions were screened and separated by HPLC. After enzymatic hydrolysis, the liquid chromatography-mass spectrometry/mass spectrometry(LC-MS/MS) was analyzed, and Bio Work 3.2 was used to analyze the data. The expression of serum egg white markers in the gastric cancer group and the control group was detected by immunoblotting. The results showed that three different protein peaks were found in the serum of gastric cancer group, which were m/z 5 905.6, m/z 6 638.5 and m/z 8 710.3 respectively. Compared with the control group, the expression signal of m/z 5 905.6 peak in gastric cancer group was enhanced((7.95±3.32) for gastric cancer group and(0.77±0.21) in control group), while the expression signal of m/z 6 638.5 wasp(gastric cancer group was(5.78 ±2.55), control group was(16.45 ±4.32)) and m/z 8 710.3 peak(gastric cancer group(0.94±0.19), control group was(2.23±0.87)). The specific protein markers in the serum of m/z 5 905.6, m/z 6 638.5 and m/z 8 710.3 were used to determine the specific protein markers in the gastric cancer patients and the control groups. The detection rates of gastric cancer group and control group were 93.3%and 95% respectively. The results of identification showed that m/z 5 905.6, m/z 6 638.5 and m/z 8 710.3 three wasps were fibrinogen alpha chain(fibrinogen alpha chain, FGA), apolipoprotein A-Ⅱ(apolipoprotein A-Ⅱ, Apo A-Ⅱ) and apolipoprotein. Western blotting results showed that the expression of FGA in gastric cancer patients increased significantly(p<0.05), and the expression of Apo A-II and Apo C-Ⅰ decreased significantly(p<0.05). The research data show that the expression of three serum protein markers in FGA, Apo A-Ⅱ and Apo C-Ⅰ in gastric cancer is related to the progression of gastric cancer, and has potential diagnostic value in clinical application.