Objective: To develop a sensitive method to detect minimal residual disease and to elucidate the significance of bcl-2 gene rearrangement in diagnosis and treatment of malignant lymphoma. Methods: Using polymerase cha...Objective: To develop a sensitive method to detect minimal residual disease and to elucidate the significance of bcl-2 gene rearrangement in diagnosis and treatment of malignant lymphoma. Methods: Using polymerase chain reaction (PCR) to detect bcl-2 gene rearrangement and using serial dilution method to define the sensitivity of PCR. Results: In 9 different malignant lymphoma cell lines, Su-DHL-4 and Su-DHL-6 were shown bcl-2(MBR)/JH rearrangement, the sensitivity of PCR was 1:105. In 16 patients with follicular lymphoma, the peripheral blood and bone marrow were PCR positive in 4 cases both at initial diagnosis and after complete remission. Conclusion: Detection of bcl-2 gene rearrangement by PCR provides a sensitive and specific assay of minimal residual disease. It is helpful to improve staging of disease, prognosis and evaluation of the treatment results.展开更多
The rearrangement of Bcl-1 gene (Bcl-1/IgH rearrangement) and expression of cyclin D1 in multiple myeloma (MM) precursor cells were studied and the role of cyclin D1 in the pathogenesis of MM was investigated. The BCL...The rearrangement of Bcl-1 gene (Bcl-1/IgH rearrangement) and expression of cyclin D1 in multiple myeloma (MM) precursor cells were studied and the role of cyclin D1 in the pathogenesis of MM was investigated. The BCL-1 rearrangement and cyclin D1 protein expression in 15 cases of MM were detected. By using hemi-nested polymerase chain reaction (PCR) the genomic DNA from fresh peripheral blood and bone marrow was amplified and the expression of cyclin D1 in the smears was detected by using immunohistochemical method. Ten volunteer with normal bone marrow served as control group. The results showed Bcl-1 rearrangement was detectable in 3/15 (20 %) MM patients and cyclin D, expression in 4/15 (27 % ) MM patients. BeLl-1 rearrangement and cyclin D1 protein expression were also detected in MM precursor cells. No overexpression of cyclin D1 or the rearrangement of the BeL-1 gene was found in the 10 volunteers. It was concluded that Bel-1 rearrangement and cyclin D1 protein overexpression were detected in MM precursor cells, speculating that overexpression of cyclin D1 protein may play an initial (critical) role in the pathogenesis of MM.展开更多
目的探讨Bcl-2在弥漫性大B细胞淋巴瘤（diffuse large B—cell lymphoma，DLBCL）不同类型中的表达及意义。方法应用免疫组织化学S—P法检测60例弥漫性大B细胞淋巴瘤标本中Bcl-2蛋白的表达水平；同时行RT—PCR检测外周血Bcl-2／IgH的表...目的探讨Bcl-2在弥漫性大B细胞淋巴瘤（diffuse large B—cell lymphoma，DLBCL）不同类型中的表达及意义。方法应用免疫组织化学S—P法检测60例弥漫性大B细胞淋巴瘤标本中Bcl-2蛋白的表达水平；同时行RT—PCR检测外周血Bcl-2／IgH的表达水平。结果60例DLBCL中，Bcl-2阳性率分别为61．67％（37／60）；Bcl-2／IgH的阳性表达12例（12／60）。结论Bcl-2有望成为DLBCL亚分类的指标。展开更多
目的寻找一种敏感、特异的方法检测弥漫性大B细胞淋巴瘤(DLBCL)bcl-2/IgH基因重排,并通过测定产物的序列,以了解所建方法的可靠性。方法运用专业引物设计软件设计bcl-2/IgH基因重排半巢式PCR引物,对52例经临床病理确诊的DLBCL石蜡包埋...目的寻找一种敏感、特异的方法检测弥漫性大B细胞淋巴瘤(DLBCL)bcl-2/IgH基因重排,并通过测定产物的序列,以了解所建方法的可靠性。方法运用专业引物设计软件设计bcl-2/IgH基因重排半巢式PCR引物,对52例经临床病理确诊的DLBCL石蜡包埋组织及10例慢性扁桃体炎患者的新鲜扁桃体组织通过半巢式递降温度梯度PCR(touch down PCR)扩增,检测bcl-2/IgH基因重排,并对其产物进行克隆和序列分析。结果通过一步法检测到bcl-2/IgH基因重排8例,其中DLBCL6例,新鲜扁桃体组织2例;进行第2次半巢式PCR时仅在DLBCL中发现5例阳性,新鲜扁桃体组织均阴性。将阳性产物在网上序列分析显示:一步法检测到的8例中有3例为假阳性,而半巢式PCR扩增出的5例均为bcl-2/IgH基因重排片段。3例假阳性的片段分别与人类第19号染色体BAC331191,LLNLR-245D11基因片段及1号染色体RP11-498P10基因片段同源。结论常用的检测bcl-2/IgH基因重排引物扩增结果存在一定假阳性,其机制可能是因为人类基因组中存在与常用引物同源性较高的序列。为研究设计的引物与传统的引物结合,进行半巢式PCR可以排除这种假阳性扩增,提高诊断的准确性。展开更多
文摘Objective: To develop a sensitive method to detect minimal residual disease and to elucidate the significance of bcl-2 gene rearrangement in diagnosis and treatment of malignant lymphoma. Methods: Using polymerase chain reaction (PCR) to detect bcl-2 gene rearrangement and using serial dilution method to define the sensitivity of PCR. Results: In 9 different malignant lymphoma cell lines, Su-DHL-4 and Su-DHL-6 were shown bcl-2(MBR)/JH rearrangement, the sensitivity of PCR was 1:105. In 16 patients with follicular lymphoma, the peripheral blood and bone marrow were PCR positive in 4 cases both at initial diagnosis and after complete remission. Conclusion: Detection of bcl-2 gene rearrangement by PCR provides a sensitive and specific assay of minimal residual disease. It is helpful to improve staging of disease, prognosis and evaluation of the treatment results.
文摘The rearrangement of Bcl-1 gene (Bcl-1/IgH rearrangement) and expression of cyclin D1 in multiple myeloma (MM) precursor cells were studied and the role of cyclin D1 in the pathogenesis of MM was investigated. The BCL-1 rearrangement and cyclin D1 protein expression in 15 cases of MM were detected. By using hemi-nested polymerase chain reaction (PCR) the genomic DNA from fresh peripheral blood and bone marrow was amplified and the expression of cyclin D1 in the smears was detected by using immunohistochemical method. Ten volunteer with normal bone marrow served as control group. The results showed Bcl-1 rearrangement was detectable in 3/15 (20 %) MM patients and cyclin D, expression in 4/15 (27 % ) MM patients. BeLl-1 rearrangement and cyclin D1 protein expression were also detected in MM precursor cells. No overexpression of cyclin D1 or the rearrangement of the BeL-1 gene was found in the 10 volunteers. It was concluded that Bel-1 rearrangement and cyclin D1 protein overexpression were detected in MM precursor cells, speculating that overexpression of cyclin D1 protein may play an initial (critical) role in the pathogenesis of MM.
文摘目的探讨Bcl-2在弥漫性大B细胞淋巴瘤（diffuse large B—cell lymphoma，DLBCL）不同类型中的表达及意义。方法应用免疫组织化学S—P法检测60例弥漫性大B细胞淋巴瘤标本中Bcl-2蛋白的表达水平；同时行RT—PCR检测外周血Bcl-2／IgH的表达水平。结果60例DLBCL中，Bcl-2阳性率分别为61．67％（37／60）；Bcl-2／IgH的阳性表达12例（12／60）。结论Bcl-2有望成为DLBCL亚分类的指标。
文摘目的寻找一种敏感、特异的方法检测弥漫性大B细胞淋巴瘤(DLBCL)bcl-2/IgH基因重排,并通过测定产物的序列,以了解所建方法的可靠性。方法运用专业引物设计软件设计bcl-2/IgH基因重排半巢式PCR引物,对52例经临床病理确诊的DLBCL石蜡包埋组织及10例慢性扁桃体炎患者的新鲜扁桃体组织通过半巢式递降温度梯度PCR(touch down PCR)扩增,检测bcl-2/IgH基因重排,并对其产物进行克隆和序列分析。结果通过一步法检测到bcl-2/IgH基因重排8例,其中DLBCL6例,新鲜扁桃体组织2例;进行第2次半巢式PCR时仅在DLBCL中发现5例阳性,新鲜扁桃体组织均阴性。将阳性产物在网上序列分析显示:一步法检测到的8例中有3例为假阳性,而半巢式PCR扩增出的5例均为bcl-2/IgH基因重排片段。3例假阳性的片段分别与人类第19号染色体BAC331191,LLNLR-245D11基因片段及1号染色体RP11-498P10基因片段同源。结论常用的检测bcl-2/IgH基因重排引物扩增结果存在一定假阳性,其机制可能是因为人类基因组中存在与常用引物同源性较高的序列。为研究设计的引物与传统的引物结合,进行半巢式PCR可以排除这种假阳性扩增,提高诊断的准确性。
