摘要
目的探讨miR-15a和miR-16-1寡核苷酸能否增强Raji细胞对阿糖胞苷(cytarabine,Ara-C)的敏感性。方法将化学合成的miR-15a和miR-16-1寡核苷酸利用脂质体2000转染入Raji细胞后,联合Ara-C,CCK8法检测Ara-C的IC50值变化;锥虫蓝细胞计数法检测细胞增殖活性;Hoechst染色观察细胞的凋亡形态;流式细胞仪AnnexinⅤ/PI双染法测凋亡率。结果miR-15a和miR-16-1寡核苷酸转染Raji细胞后,再加入Ara-C,24h Ara-C的IC50值分别为10.41和10.86,明显低于单用Ara-C组(15.43)和随机序列联用Ara-C组(14.92,P<0.05)。锥虫蓝拒染法结果显示,转染后各时间点miR-15a/miR-16-1+Ara-C组较单用miR-15a/miR-16-1组、单用Ara-C组及随机序列+Ara-C组明显抑制了Raji细胞的生长,Hoechst染色可见大量凋亡细胞;流式细胞仪AnnexinⅤ/PI双染色法检测显示,miR-15a+Ara-C组早期凋亡率和晚期凋亡率分别为20.93%和25.27%,miR-16-1+Ara-C组早期凋亡率和晚期凋亡率分别为20.69%和23.13%,均明显高于miR-15a组、miR-16-1组、Ara-C组及随机序列+Ara-C组(P<0.05),后四者的早期凋亡率分别为6.99%、4.73%、10.88%和14.39%,晚期凋亡率分别为10.08%、10.64%、11.83%和11.93%。结论miR-15a和miR-16-1寡核苷酸可增强Raji细胞对Ara-C的敏感性。
Objective To study whether miR-15a and miR-16-1 can enhance the sensitivity of Raji cells to cytarabine（Ara-C）.Methods MiR-15a and miR-16-1 oligonucleotides were transfected into Raji cells with LipofectamineTM 2000,and then the cells were treated with Ara-C.The IC50 values of Ara-C was detected by CCK8 assay.The growth of Raji cells was measured by trypan blue dye exclusion method.The apoptotic cells were observed by Hoechst dyeing;AnnexinⅤ/PI double dyeing and glow cytometry（FCM） were used to examine the cell apoptotic rate.Results After transfection of miR-15a or miR-16-1 into Raji cells,the IC50 values of Ara-C were 10.41 and 10.86,respectively,which were significantly lower than that of the untransfected group（15.43）and scrambled oligonucleotides（SODN）transfection group（14.92,P0.05）.Trypan blue dye exclusion assay showed that miR-15a/miR-16-1 transfection group had obviously decreased the cell growth compared to miR-15a,miR-16-1 group,untransfected group and SODN transfected group;Hoechst dyeing demonstrated plenty of apoptotic cells.AnnexinⅤ/PI double dyeing assays by FCM indicated that the cell apoptotic rates in earlier period and late period were 20.93% and 25.27% in the miR-15a＋Ara-C group,and 20.69% and 23.13% in the miR-16-1＋Ara-C group,which were obviously higher than those in miR-15a group（6.99%,10.08%）,miR-16-1 group（4.73%,10.64%）,Ara-C group（10.88%,11.83%） and control group（14.39%,11.93%）.Conclusion MiR-15a and miR-16-1 oligonucleotides can enhance the sensitivity of Raji cells to Ara-C.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2010年第3期274-277,共4页
Academic Journal of Second Military Medical University
基金
广东省自然科学基金(04010446)
国务院侨办重点学科建设基金(51205002)~~
