逆转录聚合酶链反应检测石蜡包埋组织中细胞周期蛋白D1 mRNA对套细胞淋巴瘤的诊断价值  

Detection of cyclin D1 mRNA by reverse transcription-polymerase chain reaction in Paraffinembedded tissues and its diagnostic significance for mantle cell lymphoma

作　　者：水若鸿[1] 魏永昆[1] 孙孟红[1] 王坚[1] 施达仁[1] 张太明[1] 金爱萍[1] 朱雄增[1] 

机构地区：[1]复旦大学附属肿瘤医院病理科,上海200032

出　　处：《中华病理学杂志》2005年第8期514-518,共5页Chinese Journal of Pathology

摘　　要：目的探讨用逆转录聚合酶链反应(RTPCR)法和竞争性RTPCR法检测套细胞淋巴瘤(MCL)石蜡包埋组织中细胞周期蛋白D1(cyclinD1)蛋白和mRNA在常规病理工作中的可行性及其诊断和鉴别诊断价值。方法收集淋巴结内MCL38例、对照组包括结内小B细胞淋巴瘤58例(B小淋巴细胞性淋巴瘤14例,淋巴浆细胞性淋巴瘤3例,滤泡性淋巴34例,淋巴结边缘区B细胞淋巴瘤7例)和淋巴结反应性增生病例20例,用免疫组织化学EnVision法和RTPCR法、竞争性RTPCR法检测cyclinD1蛋白及其mRNA的表达,以看家基因PGK作为内对照检测RNA。结果(1)38例结内MCL中,cyclinD1蛋白阳性率为71.1%(27/38),对照组均为阴性。(2)116例标本中,可检出内对照PGK基因mRNA表达103例(88.8%)。38例MCL中PGK阳性36例(94.7%)。(3)38例结内MCL中,34例可检出cyclinD1mRNA表达,去除PGK和cyclinD1mRNA均阴性的2例,MCL中cyclinD1mRNA表达的阳性率为94.4%(34/36)。对照组中B小淋巴细胞性淋巴瘤1例检出cyclinD1mRNA表达,其余病例均未检出cyclinD1mRNA表达。PCR结果全部经测序证实。(4)用竞争性RTPCR,38例结内MCL中27例可检出cyclinD1mRNA高表达,去除2例PGK也为阴性的病例,MCL中cyclinD1mRNA高表达率为75.0%(27/36)。对照组小B细胞恶性淋巴瘤及淋巴结反应性增生无1例有cyclinD1mRNA高表达。结论RTPCR方法和竞争性RTPCR方法可在石蜡包埋组织中检测cyclinD1mRNA的表达,均可用于MCL的诊断。Objective To investigate the feasibility of detecting cyclin D1 mRNA in paraffin-embedded tissues by reverse transcriptase polymerase chain reaction （RT-PCR） and competitive RT-PCR and its diagnostic and differential diagnostic significance for mantle cell lymphoma （MCL）. Methods Paraffin-embedded samples of 36 cases of MCL, 71 cases of other small B-cell lymphomas and 20 cases of lymphoid reactive hyperplasia as control group were retrieved from archival materials. Cyclin D1 protein and its mRNA was detected by EnVision and RT-PCR and competitive RT-PCR in all samples. House-keeping gene PGK was choosen as internal control. Results （ 1 ） Cyclin D1 protein was expressed in 27 of the 38 MCL （71.1%）. No cyclin D1 expression was found in the control group. （2） PGK was detected in 103 of the 116 cases （88. 8% ） and also detected in 34 of 36 MCL cases （94.7%）. （3） cyclin D1 mRNA was detected in 34 nodal mantle cell lymphoma cases by RT-PCR in paraffin-embedded tissues. The positive rate of cyclin D1 mRNA was 94. 4% in mantle cell lymphomas after exclusion of the 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia, except 1 case of B-SLL. Sequencing analysis showed that sequences were identical to cyclin D1. （4） Cyclin D1 mRNA overexpression was detected in 27 cases of nodal mantle cell lymphoma by competitive RT-PCR in paraffin-embedded tissues. The positive rate of cyclin D1 mRNA overexpression was 75.0% in mantle cell lymphomas after exclusion of 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA overexpression was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia. Conclusion RT-PCR and competitive RT-PCR detection of cyclin D1 mRNA overexpression could be used for the diagnosis and differential diagnosis of mantle cell lymphoma in paraffin-embedded blocks.

关 键 词：淋巴瘤 膜细胞 细胞周期蛋白D1 逆转录聚合酶链反应 诊断 鉴别 基因MRNA表达 聚合酶链反应检测 套细胞淋巴瘤 石蜡包埋组织 

分 类 号：R733.1[医药卫生—肿瘤]