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目的分析单个霍奇金淋巴瘤细胞IκBα基因第5外显子的碱基序列,探讨IκBα基因突变与霍奇金淋巴瘤的关系。方法用免疫组织化学EnVision法检测7例霍奇金淋巴瘤CD30抗原,采用激光显微切割技术捕获CD30阳性表达的单个霍奇金淋巴瘤细胞和其周围反应性增生的淋巴细胞,以这些标本的基因组DNA为模板,用半巢式聚合酶链反应(PCR)方法扩增IκBα基因的第5外显子,PCR产物经纯化后直接测序。结果分析测序图发现3例霍奇金淋巴瘤细胞IκBα第5外显子出现点突变,均形成TGA终止密码子。结论霍奇金淋巴瘤中IκBα基因第5外显子出现点突变,可能是霍奇金淋巴瘤恶性转化过程中除EB病毒感染外另一重要分子机制。
Objective To analyze the base sequence of exon 5 of IκBα gene in single Hodgkin lymphoma and to explore the relationship between IκBα gene mutation and Hodgkin ’s lymphoma. Methods Seven cases of Hodgkin ’s lymphoma CD30 antigen were detected by immunohistochemistry EnVision method. Single Hodgkin’ s lymphoma cells with CD30 positive expression and lymphocytes with reactive hyperplasia were captured by laser microdissection. Genomic DNA was used as a template to amplify the fifth exon of IκBα gene by semi-nested polymerase chain reaction (PCR). The PCR products were directly sequenced after purification. Results Analysis of sequence map found point mutations in IκBα exon 5 of three Hodgkin lymphoma cells, all forming TGA stop codons. Conclusions Point mutation of exon 5 of IκBα gene in Hodgkin ’s lymphoma may be another important molecular mechanism in addition to Epstein - Barr virus infection during the malignant transformation of Hodgkin’ s lymphoma.