hsa-miR-150再表达对Burkitt淋巴瘤增殖与凋亡的影响

Effects of re-expression of hsa-miR-150 on proliferation and apoptosis in Burkitt's lymphomas

目的探讨hsa-miR-150在正常人B淋巴细胞群及人Burkitt淋巴瘤(Burkitt’s lymphoma,BL)中的表达,分析hsa-miR-150再表达对人BL细胞株Daudi及Raji体外增殖及凋亡的影响。方法采用流式细胞技术(flow cytometry,FCM)从扁桃体炎标本中分选出正常B淋巴细胞群,应用qRT-PCR技术检测hsa-miR-150在细胞群中的表达,同时检测hsa-miR-150在BL细胞株Daudi及Raji中的表达;将载有hsa-miR-150的慢病毒稳定转染Daudi及Raji,采用FCM、MTT及IP染色法观察hsa-miR-150再表达对Daudi及Raji细胞增殖与凋亡的影响。结果 FCM从扁桃体组织中分选出4群正常的B淋巴细胞群,分别为nave细胞(NC)、中心母细胞(CB)、中心细胞(CC)、记忆B细胞(MC),hsa-miR-150在NC、CB、CC、MC中的表达呈动态性,相对于NC,CB及CC低表达hsa-miR-150(P<0.001)。与正常B淋巴细胞相比,hsa-miR-150在Daudi及Raji中明显低表达(P<0.001)。与对照组相比,实验组细胞增殖明显受抑制(P<0.001),凋亡明显增加(P<0.001)。结论 hsa-miR-150对正常B淋巴细胞的分化具有重要作用,与BL的发生、发展可能存在一定关系;hsa-miR-150再表达可抑制Daudi及Raji细胞增殖,诱导其凋亡,为后续诱导分化治疗研究打下基础。

Purpose To investigate expression of human small non-code RNA hsa-miR-150 in normal human B lymphocyte subsets and Burkitt＇ s lymphomas （ BL）, and to analyze effect of re-expression of hsa-miR-150 on proliferation and apoptosis in human BL cell lines Daudi and Raji. Methods Human normal B lymphocyte subsets were selected by flow cytometry （FCM） from human tonsils, and real time-PCR （qRT-PCR） was employed to detect expression of hsa-miR-150 in the subsets and Daudi and Raji; lentivirus, FCM, MTT and IP staining were used to analyze the effect of re-expression of hsa-miR-150 on Daudi and Raji cells. Results Naive cells （ NC）, follicular centroblasts （CB）, follicular centrocytes （CC） and memory B cells （MC） were selected, Hsa-miR-150 expression in NC, CB, CC and MC was dynamically changed and it was significantly decreased in CB and CC compared with NC （P 〈 0. 001 ）, it was also lower in Daudi and Raji than that of normal B cells （P 〈 0. 001 ）. Proliferation was significantly suppressed and apoptosis was induced in experimental groups compared with control groups （P 〈 0. 001 ）. Conclusion Dynamic expression of hsa-miR-150 in normal B lymphocyte subsets may play an important role in differentiation of human normal B cells, dysregulated expression of hsa-miR-150 in human BL may result in development of B cell lymphomas. Re-expression of hsa-miR-150 suppresses proliferation and induces apoptosis of Daudi and Raji cells. Hsa-miR-150 may serve as a bona fide therapeutic target.