mir-15a和mir-16-1诱导人淋巴瘤Raji细胞株的凋亡  

The effect of mir-15a and mir-16-1 on the apoptosis of Raji lymphomatic cells

作　　者：谌琴[1] 何冬梅[1] 张洹[1] 郭敏[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：《暨南大学学报（自然科学与医学版）》2008年第6期567-572,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省自然科学基金项目(04010446)

摘　　要：目的:探讨mir-15a和mir-16-1诱导人淋巴瘤Raji细胞株的凋亡。方法:利用Oligofectamine 2000脂质体法将化学合成的mir-15a和mir-16-1寡核苷酸,随机序列,分别转染Raji细胞。采用间接免疫荧光及流式细胞仪检测Bcl-2蛋白表达情况,半定量RT-PCR检测Bcl-2 mRNA表达水平,台盼蓝拒染法和CCK8法检测mir-15a和mir-16-1对Raji细胞的生长抑制作用,Hoechst染色观察细胞凋亡形态,流式细胞仪—AnnexinV/PI双染色法检测细胞凋亡率。结果:间接免疫荧光法显示Raji细胞经转染mir-15a和mir-16-1寡核苷酸48 h后,Bcl-2蛋白的表达量降低,与空白对照组和随机序列组有显著性差异(P<0.05);RT-PCR法显示各组间Bcl-2 mRNA的表达无显著性差异;台盼蓝拒染法和CCK8法显示,转染后各时间点mir-15a和mir-16-1寡核苷酸组Raji细胞的生长受到抑制,Hoechst染色可见明显凋亡细胞,流式细胞仪—AnnexinV/PI双染色法检测显示,mir-15a组早期凋亡率和晚期凋亡率分别为9.74%和9.65%,mir-16-1组早期凋亡率和晚期凋亡率分别为9.70%和9.34%,明显高于空白对照组和随机对照组。结论:mir-15a和mir-16-1可诱导淋巴瘤细胞凋亡。Aim：To explore the apotosis effects of mir-15a and mir-16-1 in inducing Raji cell line. Methods： mir-15a and mir-16-1 oligonucleotides,control group were all transfected into Raji cells with lipofectamine 2000. Bcl-2 protein expression of every group cells was detected by indirect immunofluorescence ;Semi-quantitative RT-PCR detected the expression level of Bcl-2 mRNA; The growth inhibitory effect of Raji cells was measured by trypan blue dye exclusion method and CCK8 assay. The apoptotie ceils were observed by Hoechst Method; Flow Cytometry （FCM）-AnnexinV/PIdouble dyeing method was used to detect the cell apoptotic rate. Results： After Raji cells were transfected for 48 h, Bcl-2 protein expression levels of mir-15a group and mir-16-1 group cells obviously decreased, The above two groups had obviously difference as compared with the control group and blank group （ P 〈 0. 05 ）. The expression of Bcl-2 mRNA among the four groups had no obviously difference,. Trypan blue dye exclusion method and CCK8 assay showed that transfection of mir-15a and mir-16-1 decreased the cell growth at 24, 48 and 72 h post-transfection, apoptotic cells can be seen with Hoechst Method at 48 h after being transfected. FCM assays indicated that the cell apoptotic rates in earlier period and advanced stage of mir- 15a group were 9.74% and 9.65% ,and mir-16-1 group were 9.70% and 9. 34% ,which were obviously higher than blank group and control group. Conclusion： mir-15a and mir-16-1 can induce apoptosis of lymphomatic ceils.

关 键 词：微RNA 淋巴瘤 微RNA-15 微RNA-16 细胞凋亡 

分 类 号：R733.4[医药卫生—肿瘤] R394.3[医药卫生—临床医学]