Proteomic analysis to identify biomarker proteins in pancreatic ductal adenocarcinoma
Proteomic analysis to identify biomarker proteins in pancreatic ductal adenocarcinoma
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DOI：
10.1111/j.1445-2197.2008.04429.x
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发表时间：
2008-04-01
影响因子：
1.7
通讯作者：
Bae, Chang Dae
中科院分区：
文献类型：
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作者：
Chung, Jun Chul;Oh, Mi Jung;Bae, Chang Dae
背景：胰腺导管腺癌（PDAC）是韩国第五大常见的癌症死亡原因。PDAC很难在早期诊断，甚至更难治愈。因此，迫切需要鉴定用于早期诊断和有效治疗的分子靶点。本研究的目的是使用蛋白质组学分析鉴定PDAC差异表达的生物标志物蛋白，使用蛋白质印迹分析验证鉴定的与癌发生相关的生物标志物蛋白，并评估影响候选生物标志物蛋白表达的临床因素。在本研究中，我们对10对PDAC标本与匹配的邻近正常组织进行了蛋白质组学分析，以阐明蛋白质表达的不同模式。采用高分辨双向聚丙烯酰胺凝胶电泳（2D PAGE）分离蛋白质，基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）鉴定差异表达蛋白质。使用蛋白质印迹分析进一步验证与癌发生相关的候选生物标志物蛋白的差异表达。标准的统计分析进行了尝试，以建立一个临床变量和候选生物标志物protein.Results的表达之间的相关性：PDAC和相邻的正常组织的分析显示，可重复相似的蛋白质组模式为每组。通过银染凝胶从PDAC和正常组织中观察到约700个斑点。差异表达的蛋白质点进行凝胶消化和MALDI-TOF MS鉴定。25个蛋白质被鉴定，其中5个蛋白质（半乳糖凝集素-1，烯醇化酶-2，α-1-抗胰蛋白酶，N-myc相互作用，过氧化物酶氧还蛋白-4）先前被报道为在mRNA水平或蛋白质水平在人类癌症中差异表达。选择这五种蛋白质作为与致癌相关的候选生物标志物蛋白。通过蛋白质印迹分析进一步验证了这些蛋白质。在候选生物标志物蛋白中，半乳糖凝集素-1表达与组织学高度相关（P = 0.019），T分期（P = 0.047），N期（P = 0.033）和美国癌症联合委员会分期（P = 0.011）。结论：通过蛋白质组学分析鉴定了PDAC中差异表达的25个蛋白质，并验证了5个与癌发生相关的蛋白质。蛋白质印迹分析。galectin-1表达与肿瘤组织学和分期高度相关。
Background: Pancreatic ductal adenocarcinoma (PDAC) is the fifth most common cause of death from cancer in Korea. PDAC is difficult to diagnose at an early stage and even more difficult to cure. Thus, there is an urgent need to identify molecular targets for early diagnosis and effective treatment. The objectives of this study were to identify differentially expressed biomarker proteins of PDAC using proteomic analysis, to validate the identified biomarker proteins associated with carcinogenesis using western blot analysis and to evaluate clinical factors influencing expression of candidate biomarker proteins.Methods: In the present study, we carried out proteomic analysis in 10 pairs of PDAC specimens with matching adjacent normal tissues to clarify the different patterns of protein expression. The proteins were separated by high-resolution 2-D polyacrylamide gel electrophoresis (2D PAGE) and the differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Differential expression of candidate biomarker proteins associated with carcinogenesis was further validated using western blot analysis. Standard statistical analysis was carried out in an attempt to establish a correlation between clinical variables and expression of candidate biomarker proteins.Results: Analysis of PDAC and the adjacent normal tissues showed reproducibly similar proteomic patterns for each group. Approximately 700 spots each were seen by silver-stained gels from both PDAC and normal tissues. Differentially expressed protein spots were gel digested and identified by MALDI-TOF MS. Twenty-five proteins were identified, of which five proteins (galectin-1, enolase-2, alpha-1-antitrypsin, N-myc interactor, peroxiredoxin-4) were previously reported as being differentially expressed either at the mRNA level or protein level in human cancer. The five proteins were selected for candidate biomarker proteins related to carcinogenesis. These proteins were further validated by western blot analysis. Among the candidate biomarker proteins, galectin-1 expression was highly correlated to histology (P = 0.019), T stage (P = 0.047), N stage (P = 0.033) and American Joint Committee on Cancer stage (P = 0.011).Conclusion: Differentially expressed 25 proteins in PDAC were identified using proteomic analysis and five proteins related to carcinogenesis were validated by western blot analysis. galectin-1 expression was highly correlated to tumour histology and stage.