Article Abstract

Effect of miRNA-19a antisense oligonucleotide and Ara-C on the proliferation and apoptosis of HL60 cells

Authors: Shijie Bao, Dongmei He, Jinrong Zeng, Yanrong Zhang, Shengting Chen

Abstract

Background: This study aimed to investigate the effects of microRNA19a (miR-19a) antisense oligonucleotide (ASODN) on the proliferation and apoptosis of acute myeloid leukemia cells (HL60).
Methods: In experiment 1, HL60 cells were divided into the blank control group, the blank transfection group, the scrambled oligonucleotide (SODN) group and the ASODN group. MiR-19a ASODN and SODN were independently transferred into HL60 cells. The miR-19a expression was detected by real-time quantitative RT-PCR (qRT-PCR) after 48-h and 72-h transfection; CCK8 assay was used to detect the proliferation inhibition rate at 48 and 72 h; Hoechst 33258 staining was performed to examine apoptotic cells at 48 h; the apoptosis rate was detected by flow cytometry after AnnexinV/PI staining at 48 and 72 h; the protein expression of E2F1 and Bim was detected by Western blotting at 48 h. In experiment 2, cells were divided into the Ara-C group, the SODN + Ara-C group and the ASODN + Ara-C group. The cell proliferation inhibition rate at 48 and 72 h and apoptosis rate at 72 h were assessed as mentioned above.
Results: MiR-19a expression in the miR-19a ASODN group was lower than in the SODN group and the blank control group (P<0.05). MiR-19a ASODN significantly inhibited the growth of HL60 cells (P<0.05) and increased their apoptosis, and the apoptosis rate peaked at 48 h. The protein expression of E2F1 and Bim in the ASODN group was higher than in the blank control group, blank transfection group and SODN group. In addition, Ara-C further inhibited the growth and induced the apoptosis of miR-19a ASODN-transfected cells (P<0.05) in a time dependent manner. The growth inhibition rate and apoptosis rate in the ASODN + Ara-C group were higher than the sum of those in both Ara-C group and ASODN group.
Conclusions: miRNA-19a ASODN can inhibit the proliferation and induce apoptosis of HL60 cells and may exert synergistic effects with Ara-C on HL60 cells.