原位PCR方法在霍奇金淋巴瘤研究中的应用

In Situ PCR in the Study of Hodgkin's Lymphoma

目的1.探索在石蜡切片上进行直接法原位PCR的可行性。2.探讨霍奇金淋巴瘤(Hodgkin’sLym-phoma,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,采用该方法对3例福尔马林固定石蜡包埋的HL的H/R-S细胞IgH基因重排情况进行检测,综合分析结果。结果2例结节硬化型HL(NSHL)中,1例有VH2、VH3、FR3基因重排,1例有VH1、VH基因重排;1例混合细胞型HL(MCHL)的H/R-S细胞有VH2、VH3、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.在石蜡切片上行直接法原位PCR是可行和有效的。2.VH1、VH2、VH3、FR2a、FR3基因重排的出现,提示NSHL和MCHL的H/R-S细胞来源于不同分化阶段的B细胞,NSHLa同时出现FR1、FR3基因重排;MCHL同时出现FR1、FR2基因重排;提示NSHL和MCHL的H/R-S细胞可能为多克隆性增生。背景的部分小淋巴细胞核内也可观察到阳性信号,提示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： 1.To investigate the feasibility to proceed in situ the detection of DNA after PCR in paraffin section; 2. To investigate the origin and lineage of the H/R - S cells from HL. Methods ： Detect the DNA of the H/R - S cells after in situ PCR in Hodgkin＇s Lymphoma from three samples of paraffin section by Bio - labeled heavy chain family - specific primers. Results： 1. VH2, VH3 and FR3 gene rearrangements and VH1, VH3 gene rearrangements were respectively observed in two cases of nodular sclerosis HI, （NSHL）. 2.VH2, VH3 and FR2a gene rearrangements were revealed in one case of mixed cellarity HL （MCHL）. Positive signal was found in the nuclei of surrounding small lymphocyte. Conclusions： 1. It was feasible and validable to proceed in situ the detection of DNA after PCR in paraffin section. 2.The appearance of VHI, VH2, VH3, FR2a, FR3 gene rearrangements suggested that the H/R - S cells from classical HL （NSHIL and MCHL） originated from B lineage cells at various stages of differentiation. FR1, FR2 and FR3 gene rearrangements were demonstrated in different H/R - S cells in the same section, which suggested that H/R- S cells of NSHL and MCHL might originate from different clone of B cell, The H/R- S cells and the surrrounding background cells might originate from the same B cell clone.