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目的 :寻找一种敏感、特异而又快捷的方法检测外套细胞淋巴瘤中 bcl- 1/ Ig H 基因重排 ,并分析 bcl- 1/Ig H 基因重排产物的序列 ,以了解 bcl- 1/ Ig H 基因重排发生的确切机理。方法 :对 2 8例经临床确诊的外套细胞淋巴瘤的新鲜冰冻组织 ,通过半巢式 PCR检测 bcl- 1/ Ig H基因重排 ,并对 bcl- 1/ Ig H基因重排产物进行克隆和序列分析。结果 :通过半巢式 PCR测到有 bcl- 1/ Ig H基因重排者计 17/ 2 8例。而采用一步法 PCR仅 9例有此基因重排 ,两者相比 ,差异显著 (χ2 =4 .59,P<0 .0 5)。对 bcl- 1/ Ig H基因重排产物进行序列分析后发现 ,bcl- 1/ Ig H基因重排产物为 74～ 16 2 bp大小 ,融合基因连接区为 6～ 2 4 bp大小。在断裂点集中的 6 5bp范围内 ,找到 10个不同的断裂点 ,其中 5个未见文献报道。对 JH 基因序列的分析 ,发现 bcl- 1/ Ig H 基因重排时 ,JH1,JH3,JH4 ,JH5和 JH6都可能参与重排 ,其中 ,以 JH4多见 ,而 JH2则没有涉及。结论 :该半巢式 PCR方法是检测外套细胞淋巴瘤中 bcl- 1/ Ig H基因重排的一种敏感而特异的方法 ,对外套淋巴细胞淋巴瘤的明确诊断和治疗都有一定的指导意义 ,而对 bcl- 1/Ig H基因重排的序列分析 ,则将为外套细胞淋巴瘤的的发病机理研究提供理论依据。
OBJECTIVE: To search for a sensitive, specific, and rapid method to detect bcl-1/Ig H gene rearrangement in mantle cell lymphoma and analyze the sequence of the bcl-1/Ig H gene rearrangement product to understand bcl-1/ The exact mechanism of Ig H gene rearrangement occurs. Methods: The bcl-1/Ig H gene rearrangement was detected by semi-nested PCR in fresh frozen tissues of 28 patients with clinically confirmed mantle cell lymphoma, and the bcl-1/Ig H gene rearrangement products were cloned. Sequence analysis. Results : 17/28 cases of bcl-1/Ig H gene rearrangement were detected by semi-nested PCR. Only one case of this gene was rearranged by one-step PCR. The difference was significant (χ2 = 4.59, P<0.05). Sequence analysis of the bcl-1/IgH gene rearrangement product revealed that the bcl-1/IgH gene rearrangement product was 74 to 162 bp in size and the fusion gene linker region was 6 to 24 bp in size. Within the range of 65 bp in which breakpoints were concentrated, 10 different breakpoints were found, of which 5 were not reported in the literature. Analysis of the JH gene sequence revealed that JH1, JH3, JH4, JH5, and JH6 may be involved in rearrangement when the bcl-1/IgH gene is rearranged, of which JH4 is more common, while JH2 is not involved. Conclusion: This semi-nested PCR method is a sensitive and specific method for detecting bcl-1/Ig H gene rearrangement in mantle cell lymphoma. It has certain guiding significance for the diagnosis and treatment of coat lymphocytic lymphoma. The sequence analysis of the bcl-1/Ig H gene rearrangement will provide a theoretical basis for the study of the pathogenesis of mantle cell lymphoma.