B细胞淋巴瘤中18号染色体上微卫星位点与bcl-2/IgH基因重排的相关性

目的探讨B细胞淋巴瘤中18号染色体上微卫星位点D18S64、D18S61与bcl-2/IgH基因重排的相关性。方法采用巢式PCR方法对37例B细胞非霍奇金淋巴瘤bcl-2/IgH基因重排主要断裂点（MBR）进行检测,PCR-SSCP方法检测bcl-2基因内部、基因周围微卫星位点D18S64、D18S61的MSI和LOH情况。采用DNA抽提试剂盒提取石蜡包埋组织基因组DNA、全血基因组DNA-PCR扩增-变性聚丙烯酰烯酰胺凝胶垂直电泳-银染法-全自动凝胶成像系统对样本进行MSI和LOH分析。结果 37例B细胞非霍奇金淋巴瘤中,bcl-2/IgH基因重排阳性率为24.3%（9/37）。各位点MSI、LOH频率分别介于16.2%-18.9%和16.2%-21.6%。D18S61位点MSI阳性组的bcl-2基因重排阳性率均高于MSI阴性组（P〈0.05）,而D18S64位点则与之相反（P〉0.05）;D18S61位点LOH阳性组的bcl-2基因重排阳性率明显高于LOH阴性组（P〈0.05）,D18S64位点与之相反（P〉0.05）。结论 bcl-2/IgH基因重排与微卫星位点D18S61的MSI/LOH有显著相关性,表明D18S61可能是bcl-2基因周围区域与bcl-2基因关系较为密切的微卫星位点。

A study for bcl-2 gene rearrangement and microsatellite loci on 18 chromosome in B cell Non-Hodgkins lymphomas

Objective To evaluate the role of bcl-2 gene rearrangement and microsatellite instability in pathogenesis of B cell Non-Hodgkins lymphoma,and the relationship between rearrangement of bcl-2 gene and D18S64、D18S61 microsatel-lite loci on 18 chromosome.Methods Thirty-seven cases of B cell Non-Hodgkins lymphoma were studied for bcl-2 gene rearrangement in nested-PCR method;Analysis of MSI and LOH was aimed to 2 microsatellite loci（D18S61,D18S64） aro-und or in bcl-2 gene in these cases.DNA were extracted from paraffin-embedded tissues and whole human blood cells by DNA extract kits,subjected to PCR amplification,the PCR products were observed through denatured polyacrylamide gel electrophoresis and silver staining.Results Rearrangement rate of bcl-2 gene was 24.3%（9/37） in 37 non-Hodgkin lymphoma cases.Frequency of MSI and LOH varied respectively from 16.2%-18.9%,16.2%-21.6% at each locus.Analysis of correlation between bcl-2 rearrangement and MSI at different microsatellite loci showed that the bcl-2 gene rearrangement was obviously higher in the positive group of MSI at D18S61 than that of MSI negative group（P〈0.05）,which was on the contrary of D18S64（P〉0.05）.Rearrangement rate of bcl-2 at D18S61 was higher in the group positive for LOH than these in the negative group with significant difference of P〈0.05,which was on the contrary of D18S64（P〉0.05）.Conclusions Significant correlation is found between bcl-2/IgH rearrangement and MSI/LOH at microsatellite locis of D18S61,which indicated relationship between bcl-2/IgH rearrangement and D18S61 was closer than the other microsatellite loci in bcl-2 gene.