MiR-15a and miR-16-1 enhance sensitivity of Raji cells to Ara-C / 第二军医大学学报

ABSTRACT

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 Lipofectamine™ 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, P<0.05). 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.