Hsa-miR9在伯基特淋巴瘤EBV^(+/-)细胞中的差异表达及其与BCL-6调控的关系  

Expression pattern of hsa-miR-9 and its association with BCL6 in EBV positive and EBV negative Burkitt lymphoma cell lines

作　　者：代新珍[1,2] 陈少红[2] 葛娟[2] 韩西群[2] 周新华[2] 吴自勍[2] 赵彤[1,2] 

机构地区：[1]南方医科大学南方医院病理科,广东广州510515 [2]南方医科大学基础医学院病理学系,广东广州510515

出　　处：《南方医科大学学报》2013年第5期661-666,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81071941)~~

摘　　要：目的探究hsa-miR9在伯基特淋巴瘤细胞株EBV^+Raji/EBV-Ramous中差异表达及其与BCL-6的关系。方法荧光定量PCR检测hsa-miR9和BCL-6在EBV^+Raji和EBVRamous细胞中的表达;利用Oligofectamine 2000脂质体法将化学合成的hsa-miR9-inhibitor和随机序列转染高表达hsa-miR9的EBV^+Raji细胞(Raii-miR9-inhibitor组和Raji-CON组),hsa-miR9-mimics和相应的随机序列转染低表达hsa-miR9的EBVRamous细胞(Ramous-miR9-mimics组和Ramous-CON组);运用实时荧光定量PCR、Western blotting检测转染前后BCL-6的表达,AnnexinV/PI双染色法检测细胞凋亡率和观测凋亡细胞形态。结果 hsa-miR9和BCL-6在EBV^+Raji细胞的表达水平显著高于EBVRamous细胞(P<0.01);EBV^+Raji细胞经转染hsa-miR9-inhibitor 48 h后,BCL-6基因和蛋白水平的表达量均降低,而EBVRamous细胞经转染hsa-miR9-mimics 48 h后,BCL-6基因和蛋白水平的表达量均升高,与相应的随机序列组相比均有显著性差异(P<0.05);转染48 h后Raji-miR9-inhibitor组凋亡率明显低于Raji-CON组,Ramous-miR9-mimics组凋亡率明显高于Ramous-CON组,差异具有统计学意义(P<0.05);Raji-CON组和Ramous-miR9-mimics组可见明显凋亡细胞。结论 hsa-miR9正向调控伯基特淋巴瘤细胞株EBV^+Raji/EBVRamous细胞BCL-6的表达和凋亡。Objective To investigate the differential expression pattern of hsa-miR-9 between EBV-positive and -negative Burkitt lymphoma cell lines and its association with BCL-6. Methods The expression of hsa-miR9 and BCL-6 mRNAs in EBV＋ Raji and EBVRamous cells in mRNA levels were detected using fluorescence quantitative PCR （QRT-PCR）. The two cells lines were transiently transfected with hsa-mir9-inhibitor and hsa-mir9-minicsvia Oligofectamine 2000, and the changes in BCL6 expressions was detected using QRT-PCR and Western blotting. Annexin V/PI staining was used to analyze the apoptosis and morphological changes of the transfected cells. Results The expression of Hsa-miR9 and BCL-6 was significantly higher in EBV~ Raji cells than EBV- Ramous cells （P〈0.01）. BCL-6 mRNA and protein expression was reduced in EBV＋ Raji cells after transfection with hsa-miR9-inhibitor but up-regulated in EBVRamous cells transfected with hsa-miR9-minics. Flow cytometry revealed a significantly decreased apoptosis rate in EBV＋ Raji cells transfected with hsa-miR9-inhibitor but an increased rate in EBV Ramous cells transfected with hsa-miR9-minics, and the results were confirmed by microscopic observations. Conclusion Hsa-miR9 positively regulate the expression of BCL-6 and apoptosis of EBV＋ Raji cells and EBV Ramous cells.

关 键 词：伯基特淋巴瘤 hsa-miR9 BCL-6 RAJI细胞 Ramous细胞 凋亡 

分 类 号：R733.1[医药卫生—肿瘤]