5′-标记引物直接原位PCR对H/R-S细胞性质的研究

Research of H/R-S cells chacter with 5′-labeled direct in situ PCR

目的:探讨直接在石蜡切片上进行原位PCR的可行性,以此分析霍奇金淋巴瘤(hodgkinlymphoma,HL)H/R-S细胞的性质。方法:建立一种5′-标记引物直接原位PCR方法,对3例10%中性甲醛固定石蜡包埋HL的H/R-S细胞IgH基因重排情况进行检测。结果:2例结节硬化型霍奇金淋巴瘤(nodularsclerosistypehodgkinlymphoma,NSHL)H/R-S细胞中,1例有VH2、VH3和FR3基因重排,1例有VH1和VH3基因重排,1例混合细胞型霍奇金淋巴瘤(mixedcellularitytypehodgkinlymphoma,MCHL)的H/R-S细胞有VH2、VH3和FR2a基因重排。背景的部分小淋巴细胞核内也出现了阳性信号。结论:1)在石蜡切片上进行直接法原位PCR是可行和有效的。2)NSHL和MCHL的H/R-S细胞来源于不同分化阶段的B细胞,NSHL和MCHL的H/R-S细胞为多克隆性增生。周围部分背景小淋巴细胞有可能与H/R-S细胞来自同一B细胞克隆。

OBJECTIVE： To explore the feasibility to proceed in situ detection of DNA after PCR in paraffin section and investigate the origin and lineage of the H/R-S cells from classical HL. METHODS： DNA of the H/R-S cells after in situ PCR in Hodgkin lymphoma from three samples of paraffin section was detected by bio-labeled heavy chain family-specific primers. RESULTS： VH2, VH3, and FR3 gene rearrangements and VH1, VH3 gene rearrangements were respectively observed in two cases of nodular sclerosis HL （NSHL）. VH2, VH3, and FR2a gene rearrangements were revealed in one case of mixed cellarity HL （MCHL）. Positive signal was also found in the nuclei of surrounding small lymphocyte. CONCLUSIONS： 1）It is feasible and valid to proceed in situ detection of DNA after PCR in paraffin section. 2）The H/R-S cells from classical HL（NSHL and MCHL） originate from B lineage cells at various stages of differentiation. H/R-S cells of NSHL and MCHL might originate from different clones of B cell. The H/R-S cells and the surrounding background cells might originate from the same B cell clone.