同时下调长链非编码RNA PVT1和MINCR对Raji细胞增殖的影响  

Effect of down-regulating long non-coding RNA PVT1 and MINCR on proliferation of Raji cells

作　　者：郑婵丽 何冬梅[1] 刘革修[1] 李扬秋[1] 

机构地区：[1]暨南大学医学院血液病研究所,广东广州510632

出　　处：《安徽医学》2017年第5期537-541,共5页Anhui Medical Journal

基　　金：广东省科技计划项目(项目编号:2013B021800151,2015A050502029);国务院侨办重点学科建设基金项目(项目编号:51205002)

摘　　要：目的探讨同时下调长链非编码RNA(lncRNA)人浆细胞瘤转化迁移基因1(PVT1)和MINCR(MYC-induced long non-coding RNA)对淋巴瘤细胞株Raji增殖的影响及其可能机制。方法合成靶向PVT1、MINCR的小干扰RNA(siRNA)和无关对照siRNA(SC-siRNA)序列。实验分为PVT1-siRNA组、MINCR-siRNA组、PVT1-siRNA+MINCR-siRNA组、无关对照组和细胞组(未转染的Raji细胞)。将各条siRNA转入Raji细胞后,用实时定量RT-PCR方法检测MINCR RNA表达水平;分别用CCK8法、流式细胞仪、Western blot检测细胞增殖、细胞周期和c-Myc蛋白的表达情况。结果转染MINCR-siRNA后MINCR表达下调,与无关对照组和细胞组相比差异有统计学差异(P<0.05)。PVT1-siRNA联合MINCR-siRNA转染到Raji细胞后,细胞增殖明显受到抑制(P<0.05),细胞周期阻滞于G1期(P<0.05),c-Myc蛋白表达下降(P<0.05),且与单用PVT1-siRNA组、MINCR-siRNA组相比差异均具有统计学意义(P<0.05)。结论同时下调PVT1和MINCR可以增强对Raji细胞增殖的抑制作用。Objective To study the influence of down-regulating long non-coding RNA（lncRNA）plasmacytoma variant translocation 1（PVT1） and MYC-induced long noncoding RNA（MINCR）on the proliferation of lymphoma cells Raji and the probable mechanism. Methods Small interfering RNA（siRNA）targeting against PVT1 and MINCR as well as scramble control siRNA（SC-siRNA）were designed and synthesized chemically.The experiment was divided into PVT1-siRNA group, MINCR-siRNA group, PVT1-siRNA＋MINCR-siRNA group, scramble control group and untransfected cells group.Using HiperFect transfection reagent,all of the siRNAs were transfected into Raji cells.The MINCR RNA expression level was detected by real-time RT-PCR.The cell proliferation, the cell cycle and the expression of c-Myc protein were detected by CCK8 assay, Flow cytometry and Western blot, respectively. Results The expression level of MINCR RNA ofMINCR-siRNA group was significantly reduced compared with that of the scramble control group and untransfected cells group（P〈0.05）.The cell proliferation was significantly inhibited after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells （P〈0.05）,which was more obvious than that in PVT1-siRNA group and MINCR-siRNA group（P〈0.05）.Consistently,the cells were arrested in G1 phase after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells （P〈0.05）,and it was more obvious compared with PVT1-siRNA group and MINCR-siRNA group（P〈0.05）.The expression of c-Myc protein was significantly decreased after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells（P〈0.05）,which was more obvious compared with PVT1-siRNA group and MINCR-siRNA group（P〈0.05）. Conclusion Down-regulating PVT1 and MINCR can enhance the inhibition of proliferation of Raji cells.

关 键 词：长链非编码RNA 人浆细胞瘤转化迁移基因1 MINCR 小干扰RNA 细胞增殖 

分 类 号：R733.1[医药卫生—肿瘤]