目的 探讨弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma, DLBCL)中Cyclin D2的表达及其与临床病理分型及预后的关系.方法 收集79例确诊为DLBCL,每例分成两组:(1)石蜡标本制成组织芯片,应用光镜观察、免疫组化EnVision两步法检测CD3、CD20、CD10、BCL-6、MUM1、Ki-67、Cyclin D2蛋白,根据Hans分型法分型.(2)新鲜冷冻标本应用实时定量PCR(real-time quantitative PCR, qRT-PCR)法观察 Cyclin D2 mRNA在不同亚型 DLBCL中的相对表达量.应用 Kaplan-Meier 法分析Cyclin D2 mRNA相对表达量及各临床病理特征与生存期的关系.结果 79例DLBCL中,生发中心B细胞样(germinal cen-ter B-cell-like, GCB)型26例(32.9％ ),非生发中心 B细胞样(non-germinal center B-cell-like,non-GCB)型53例(67.1％ ).13例(16.5％ )表达Cyclin D2蛋白,其中2例为GCB型(15.4％ ) ,11例为non-GCB型(84.6％ ),差异有统计学意义(P＜0.05). Cyclin D2 mRNA的相对表达量在non-GCB型组显著高于GCB型组,差异有统计学意义(P＜0.05).生存分析显示高临床分期及Cyclin D2 mRNA高表达组预后差.结论 检测Cyclin D2蛋白及基因表达有助于提高DLBCL临床病理分型的准确性,并可能成为提示预后的重要指标.%Purpose To investigate the expression of Cyclin D2 in diffuse large B cell lymphoma (DLBCL) and its relationship with clinicopathological classification and prognosis. Methods (1) Immunohistochemistry (IHC) of EnVision twostep staining for CD3, CD20, CD10, BCL-6, MUM1, Ki-67 and Cyclin D2 was carried out in 79 cases of DLBCL specimens using tissue microarray constructed from tumor paraffin block.(2) The relative expression of Cyclin D2 mRNA in 79 cases of DLBCL was observed by realtime quantitative PCR (qRT-PCR) in fresh frozen tissue. The relationship between the expression of Cyclin D2 mRNA and survival rate was analyzed by Kaplan-Meier test. Results 79 patients of DLBCL were classified into26 (32.9％) case of germinal center B-cell-like (GCB) sub-types and 53 (67.1％ ) cases of nongerminal center B-cell-like(non-GCB) subtypes. Cyclin D2 protein was expressed in 2(15.4％) cases of GCB subtypes and 11 (84.6％) cases of non-GCB subtypes, respectively. The difference between the GCB subtypes and the non-GCB subtypes was statistically significant (P＜ 0.05). The expression of Cyclin D2 mRNA in non-GCB subtypes was significantly higher than that in GCB subtypes(P＜ 0.05 ). The high clinical stage and the expression of Cyclin D2 mRNA were associated with lower overall survival rate. Conclusion The detection of Cyclin D2 expression can improve the accuracy of clinicopathological classification of DLBCL. The level of Cyclin D2 mRNA may be an important reference for prognosis of DLBCL.
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