Influence of microRNA-145 on the proliferation and apoptosis of retinoblastoma Y79 cells

Abstract

Objective To observe the expression of MicroRNA 145 (miR-145) in human retinoblastoma Y79 cells and explore the effect of miR-145 on the proliferation and apoptosis of Y79 cells.Methods In this experimental study,human retinoblastoma Y79 cells were divided into four groups: miR-145 intervention group, control miRNA intervention group, empty liposome group and blank control group.miR-145 was transfected into Y79 cells by lipofection.The expression of miR-145 was detected at 48 hours after transfection by RT-PCR.Cell proliferation inhibition was measured by the Cell Counting Kit-8 (CCK-8)assay.Flow cytometery was used to examine cell cycles.Apoptosis was detected by Annexin/PI double immunofluorescence and flow cytometery.One-way ANOVA was used to analyze the overall data of every group. Results The Y79 cells were transfected by miR-145 successfully with lipofection.RT-PCR analysis showed that the expression of miR-145 in the miR-145 intervention group was significantly increased as compared to the control miRNA intervention group,empty liposome group and blank control group (79.06±3.45 vs 1.06±0.03,0.93±0.02 and 1.00±0.02;F=229.853,P＜0.05).The proliferation inhibition rate was highest in the miR-145 intervention group (21.46％) compared to the control miRNA intervention group (2.57％) and empty liposome group (3.97％)(F=34.130,P＜0.05).The cell cycle was depressed in the G1 cycle.The results of immunofluorescence and flow cytometry showed that the ratios of Annexin or Annexin/PI double positive were highest in the miR-145 intervention group,the difference was significant (F=35.434,P＜0.05).Conclusion miR-145 can inhibit proliferation and induce apoptosis in Y79 cells. Key words: Retinoblastoma; MicroRNAs; Cell proliferation; Apoptosis