全反式维甲酸诱导经典型霍奇金淋巴瘤B细胞表型  

Restoration of the lost B-cell phenotype in classical Hodgkin lymphoma cells by all trans-retinoic acid treatment

作　　者：杜静 宫凯凯 杨丽娟[1] 陈微微 罗丽卿[2] 

机构地区：[1]滨州医学院附属医院,肿瘤研究中心,山东滨州256600 [2]滨州医学院附属医院,血液科,山东演州256600

出　　处：《中国比较医学杂志》2018年第2期46-52,共7页Chinese Journal of Comparative Medicine

基　　金：山东省自然科学基金(ZR2016HB55); 烟台市科技计划(2015ZH080); 滨州医学院科研启动基金(BY2015KYQD28)

摘　　要：目的探讨全反式维甲酸(ATRA)对经典型霍奇金淋巴瘤B细胞表型的诱导作用。方法构建含有G418抗性的B细胞特异性启动子(CD19,CD79a和CD79b)表达载体,转染霍奇金淋巴瘤细胞并筛选稳定克隆。PCR扩增启动子和荧光素酶序列验证其稳定整合,敲除ABF1后检测外源B细胞启动子活性的改变以验证其功能。检测不同浓度的ATRA在不同时间点对外源B细胞启动子活性的诱导作用,通过实时定量PCR进一步证实ATRA对B细胞特异性基因转录水平的影响;检测ATRA与去甲基化药物5-Aza联合处理对B细胞标记物表达的影响;通过流式细胞染色技术仪检测ATRA对霍奇金淋巴瘤细胞表面标记物CD30表达的影响。结果 ATRA能够诱导人经典型霍奇金淋巴瘤B细胞特异性标记物CD19、CD20、CD79a和CD79b的表达,与5-Aza具有协同诱导作用,并下调细胞表面霍奇金特异性抗原CD30的表达。结论 ATRA能够诱导B细胞表型缺失的经典型霍奇金淋巴瘤向B细胞型淋巴瘤转化。Objective To investigate the induction of B-cell specific phenotype in classical Hodgkin lymphoma（ c HL） upon all-trans retinoic acid（ ATRA） incubation. Methods To construct B-cell specific promoter（ CD19,CD79 a,CD79 b） driven reporter plasmid with NEO cassette to realize stable transfection and selection of c HL reporter cells. To verify the intact integration by amplification of the promoter and luciferase sequences,and to functionally validate the B-cell specific promoter by ABF1 interference and luciferase assay. Repoter cells were incubated with various doses of ATRA and luciferase activity was detected at 24,48 and 72 hours. Reporter cells were treated alone or in combination with5-Aza and ATRA followed by luciferase assay. Endogenous B-cell specific genes（ CD19,CD20,CD79 a and CD79 b）transcription and expression levels were detected by real-time PCR and immunoblot,respectively. The expression level of CD30 antigen on Hodgkin lymphoma cell membrane upon ATRA was assessed by flow cytometry. Results ATRA treatment stimulated B-cell specific signature in c HL cells including CD19,CD79 a and CD79 b while down-regulated their CD30 expression. Conclusions ATRA induces B-cell phenotype deficient c HL cells to regain their B-cell transcriptional program while abolishes their Hodgkin-specific machinery.

关 键 词：霍奇金淋巴瘤 ATRA B细胞表型 CD19 5-AZA 

分 类 号：R-33[医药卫生]