Dai Xinzhen, Chen Shaohong, Ge Juan, Han Xiqun, Zhou Xinhua, Wu Ziqin, Zhao Tong
Department of Pathology, Southern Medical University, Guangzhou, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2013 May;33(5):661-6.
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.
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.
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.
Hsa-miR9 positively regulate the expression of BCL-6 and apoptosis of EBV(+) Raji cells and EBV(-) Ramous cells.
研究hsa-miR-9在EB病毒阳性和阴性的伯基特淋巴瘤细胞系中的差异表达模式及其与BCL-6的关系。
采用荧光定量PCR（QRT-PCR）检测EBV（+）Raji细胞和EBV（-）Ramous细胞中hsa-miR9和BCL-6 mRNA的表达水平。通过Oligofectamine 2000将hsa-mir9抑制剂和hsa-mir9模拟物分别瞬时转染至这两种细胞系，再用QRT-PCR和蛋白质免疫印迹法检测BCL6表达的变化。采用膜联蛋白V/碘化丙啶染色分析转染细胞的凋亡情况和形态变化。
EBV（+）Raji细胞中Hsa-miR9和BCL-6的表达显著高于EBV（-）Ramous细胞（P<0.01）。用hsa-miR9抑制剂转染EBV（+）Raji细胞后，BCL-6 mRNA和蛋白表达降低；而用hsa-miR9模拟物转染EBV（-）Ramous细胞后，BCL-6表达上调。流式细胞术显示，用hsa-miR9抑制剂转染的EBV（+）Raji细胞凋亡率显著降低，而用hsa-miR9模拟物转染的EBV（-）Ramous细胞凋亡率升高，显微镜观察结果证实了这一点。
Hsa-miR9正向调节EBV（+）Raji细胞和EBV（-）Ramous细胞中BCL-6的表达及细胞凋亡。
