南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (05): 661-.

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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

代新珍，陈少红，葛娟，韩西群，周新华，吴自勍，赵彤   

  • 出版日期:2013-05-20 发布日期:2013-05-20

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

  • Online:2013-05-20 Published:2013-05-20

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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EBV-Ramous 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（Raji-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EBV-Ramous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EBV-Ramous 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EBV-Ramous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/EBV-Ramous 细胞
BCL-6的表达和凋亡。

Abstract: 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
EBV-Ramous 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 EBV-Ramous 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.