Li Yin, Zhang Qin-le, Yang Xiao-li, Li Gang, He Xiao, Zhou Yi, Zang Ning, Luo Rong, Li Hong-tao, Liao Ming, Wang Yun, Zhang Xue-rong, He Min
School of Public Health, Guangxi Medical University, Nanning 530021, China.
Zhonghua Gan Zang Bing Za Zhi. 2012 Apr;20(4):275-9. doi: 10.3760/cma.j.issn.1007-3418.2012.04.010.
To determine the differentially expressed serum proteins in patients with hepatoma carcinoma and identify a putative diagnostic marker.
The isobaric tags for relative and absolute quantitation (iTRAQ) labeling method and LC-MALDI-TOF/TOF MS detection method were used to quantify serum proteins in hepatocellular carcinoma patients (n =20) and healthy individuals (n =20). Real-time reverse transcription-polymerase chain reaction was used to verify the differentially expressed proteins by analyzing the corresponding mRNA expression levels in the hepatic carcinoma and healthy hepatocyte samples, as well as in 30 pairs of patient-matched hepatic carcinoma and adjacent normal tissue samples. Western blot analysis was used to verify the protein expression in hepatic carcinoma cells.
Fifty-one proteins were significantly differentially expressed between the hepatic carcinoma group and healthy controls. The iTRAQ protein profile showed that the serum level of clusterin was significantly lower in hepatoma carcinoma patients. The mRNA level of clusterin was 20-fold lower in hepatic carcinoma cells than in healthy hepatocytes, and was 2.38-fold lower in hepatoma tissues than that in adjacent normal tissues. The clusterin protein levels were significantly lower in hepatic carcinoma cells (8.06 vs normal hepatocytes: 27.81; P less than 0.01).
The serum expression of clusterin is significantly decreased in both serum and tissues of hepatic carcinoma patients. The relationship between hepatic carcinoma and clusterin should be evaluated in future studies.
确定肝癌患者血清中差异表达的蛋白质,并鉴定一种潜在的诊断标志物。
采用相对和绝对定量的等压标签(iTRAQ)标记法和液相色谱-基质辅助激光解吸电离飞行时间串联质谱(LC-MALDI-TOF/TOF MS)检测法对肝癌患者(n = 20)和健康个体(n = 20)的血清蛋白质进行定量。通过分析肝癌和健康肝细胞样本以及30对患者匹配的肝癌和癌旁正常组织样本中相应mRNA表达水平,采用实时逆转录-聚合酶链反应验证差异表达的蛋白质。采用蛋白质印迹分析验证肝癌细胞中的蛋白质表达。
肝癌组与健康对照组之间有51种蛋白质存在显著差异表达。iTRAQ蛋白质谱显示,肝癌患者血清中簇集蛋白水平显著降低。肝癌细胞中簇集蛋白的mRNA水平比健康肝细胞低20倍,肝癌组织中比癌旁正常组织低2.38倍。肝癌细胞中簇集蛋白水平显著降低(8.06对正常肝细胞:27.81;P<0.01)。
肝癌患者血清和组织中簇集蛋白的表达均显著降低。未来研究应评估肝癌与簇集蛋白之间的关系。