5-Aza/TSA表观遗传处理诱导非霍奇金淋巴瘤B细胞表型丢失

目的:探讨5-氮杂-2’-脱氧胞苷/曲古抑菌素A(5-Aza/TSA)介导的DNA去甲基化/组蛋白乙酰化处理对B细胞来源的非霍奇金淋巴瘤B细胞表型的影响。方法:构建含有G418抗性的CD19特异性启动e GFP表达载体,转染霍奇金(阴性对照)和非霍奇金淋巴瘤细胞,并筛选目的序列稳定整合的克隆,比较CD19启动子在2组细胞中的活性。流式细胞术检测5-Aza/TSA处理对2组细胞GFP表达水平的影响。分离Eμ-myc转基因小鼠非霍奇金淋巴瘤原代细胞并鉴定其B细胞表型,通过流式细胞术检测5-Aza/TSA对其B细胞表面抗原CD19等表达水平的影响。结果:5-Aza/TSA表观遗传处理能够降低人非霍奇金淋巴瘤细胞中外源性CD19启动子的转录活性,并沉默小鼠原代非霍奇金淋巴瘤细胞表面B细胞特异性抗原的表达。结论:DNA甲基化和组蛋白去乙酰化对人类及小鼠B细胞来源的非霍奇金淋巴瘤B细胞表型稳定有重要作用。

Epigenetic modification by 5-Aza/TSA treatment induces loss of B-cell phenotype in B-cell derived non-Hodgkin lymphoma

AIM: To investigate influence of demethylation/acetylation by 5-Aza-2'-deoxycytidine/trichostatin A (5-Aza/TSA) treatment on B-cell specific phenotype of non-Hodgkin lymphoma cells. METHODS: CD19 promoter-driven reporter with NEO cassette was constructed to realize transfection and stable selection of Hodgkin and non-Hodgkin lymphoma cells. The exogenous CD19 promoter activity in both cell line clusters with and without 5-Aza/TSA treatment was detected and compared. The B-cell specific expression profiling in Eμ-myc transgenic mouse model developed lymphoma was isolated and identified. The effects of 5-Aza/TSA treatment on B-cell specific phenotype were analyzed. RESULTS: Epigenetic modification via 5-Aza/TSA repressed B-cell specific phenotype in B-cell-derived non-Hodgkin lymphoma cells. CONCLUSION: Epigenetic modification of pivotal master repressor genes plays an essential role in B-cell phenotype of both human and murine developed B-cell non-Hodgkin lymphoma cells.