Proteome analysis and tissue array for profiling protein markers associated with type B thymoma subclassification.
Proteome analysis and tissue array for profiling protein markers associated with type B thymoma subclassification.
复制标题
蛋白质组分析和组织阵列，用于分析与 B 型胸腺瘤亚分类相关的蛋白质标记。
作者：
孙强玲
背景
世界卫生组织（WHO）亚型在B型胸腺瘤中的预后相关性仍有争议。了解不同组织学类型胸腺瘤的分子特征将为B型胸腺瘤的诊断和治疗提供有意义的信息。
方法
采用双向电泳结合MALDI-TOF-MS技术对12例B型胸腺瘤组织（6例B1型，6例B2型）进行蛋白质组学分析，并对69例B型胸腺瘤组织（包括B1、B2和B3）进行组织芯片分析和免疫组化染色。采用斯皮尔曼秩相关检验（RankCorrelationTest）分析其表达与临床病理参数（如肿瘤分期或WHO分类）的关系。
结果
共鉴定出16个B1型和B2型胸腺瘤组织差异表达蛋白。使用免疫组织化学染色验证埃兹蛋白和谷胱甘肽S-转移酶pi（GSTP 1）的差异水平。Ezrin在B1型胸腺瘤和B3型胸腺瘤中的阳性表达率差异有统计学意义（Z =-2.963，P < 0.01）。Ezrin在B1-B3型的高分级肿瘤中表达。经统计学分析，B2和B3型肿瘤中GST P1的阳性表达率明显高于B1型（B2型vs. B1：Z =-2.582，P = 0.01; B3型vs. B1：Z =-4.012，P ≤ 0.001）。GST-1与B1 ~ B3肿瘤WHO分期有较强的相关性（斯皮尔曼相关系数为0.633，P ≤ 0.001）。统计学分析显示，GST 1和ezrin表达与临床分期密切相关（斯皮尔曼相关系数：ezrin：0.481，P < 0.05; GST 1：0.484，P < 0.01）。
结论
采用比较蛋白质组学方法分析B1型和B2型胸腺瘤组织中差异表达的蛋白质。蛋白质组学分析和组织芯片技术为筛选B型胸腺瘤组织学亚型的关键分子提供了潜在的工具。统计学分析ezrin和GST 1的表达，尤其是GST 1的表达，可能是一个有用的方法，B型胸腺瘤的分类。
BACKGROUND
The prognostic relevance of World Health Organization (WHO) subtypes within type B thymomas is still controversial. Understanding of the molecular characteristics of the different histologic types of thymomas will provide meaningful information for diagnosis and therapeutic management in type B thymoma.
METHODS
Proteins extracted from twelve type B thymoma tissue specimens (six type B1 and six type B2) were analyzed by two-dimensional electrophoresis (2-DE) coupled with MALDI-TOF-MS. Differentially expressed proteins were then assayed in sixty-nine type B thymoma tissues (including B1, B2 and B3) by tissue array analysis with immunohistochemistry staining. The relationship of their expression with clinicopathological parameters, such as tumor stage or WHO classification, was estimated by Spearman's Rank Correlation Test.
RESULTS
Sixteen differentially expressed proteins between type B1 and B2 thymoma tissues were identified. The differential levels of ezrin and glutathione S-transferase pi (GSTP1) were validated using immunohistochemistry staining. A statistically significant difference was observed in the positive rate of ezrin expression between type B1 thymoma and type B3 thymoma (Z = -2.963, P < 0.01). Ezrin showed a tendency to be expressed in higher classification tumors from type B1 to B3. A statistical analysis demonstrated that type B2 and B3 tumors had significantly higher positive expression of GSTP1 than the B1 group (type B2 vs. B1: Z = -2.582, P = 0.01; type B3 vs. B1: Z = -4.012, P ≤ 0.001). The results also showed a strong correlation between GSTP1 and WHO type staging of B1 to B3 tumors (Spearman's correlation coefficient: 0.633, P ≤ 0.001). Statistical analysis showed that there was close correlation between GSTP1 and ezrin expression with the clinical stage (Spearman's correlation coefficients, ezrin: 0.481, P < 0.05; GSTP1: 0.484, P < 0.01).
CONCLUSIONS
Differentially expressed proteins between type B1 and B2 thymoma tissues were analyzed by comparative proteomic analysis. The techniques of proteomic analysis and tissue array provide a potential tool for screening of key molecules in type B thymoma histological sub-classifications. The statistical analysis of ezrin and GSTP1 expression by immunohistochemistry, especially GSTP1, may be a useful approach for type B thymoma classification.