Effect of Vector-Based RNA Interference Technology on Telomerase Reverse Transcriptase Gene,Protein Expressions and Apoptosis in HL-60 Cell Line

Objective To explore the effect of RNA interference(RNAi) technology targeting telomerase reverse transcriptase(hTERT) by vector on hTERT mRNA,protein expressions and apoptosis of leukemia HL-60 cell.Methods hTERT-siRNA plasmid which had been structured was transferred into HL-60 cells by RNAi-Mate,while the empty plasmid,transferring reagent and blank cells were used as controls.Cells were collected at 24 h,72 h,120 h after transferred,and inverse transcription polymerase chain reaction was used to detect the expression of hTERT mRNA,while Western blot was used to detect the expression of hTERT protein,and flow cytometry was used to observe the apoptosis of cells.Results hTERT mRNA expression of test groups at 24 h,72 h,120 h after transferred were 0.31±0.08,0.28±0.05,and 0.32±0.06,respectively,while expression level of empty plasmid,transferring reagent and blank groups were 0.55±0.13,0.49±0.08,and 0.58±0.19,respectively.Compared with control groups,hTERT mRNA expression in test groups were significantly lower(Pa0.05);compared with blank group,hTERT mRNA in the empty plasmid and transferring reagent groups didn′t change significantly(Pa0.05).hTERT protein expression of test groups at 24 h,72 h,120 h after transferred were 0.47±0.09,0.32±0.07,and 0.51±0.08,respectively,expression level of empty plasmid,transfering reagent and blank groups were 0.75±0.11,0.76±0.15,0.73±0.14,respectively.Compared with control groups,hTERT protein in test groups was significantly lower,too(Pa0.05);but compared with blank group,hTERT protein expression in empty plasmid and transferring reagent groups had no significant change(Pa0.05).The apoptosis rate of 24 h,72 h,120 h test groups and control groups were(11.95±3.61)%,(14.61±2.97)%,(12.82±3.01)%,(4.25±2.16)%,(5.09±1.97)%,and(3.76±1.84)%,respectively.They were higher in test groups than those in control groups(Pa0.05);but compared among the test groups,there was no significant change(Pa0.05);and compared with blank group,the apoptosis rate in the empty plasmid and transferring reagent groups didn′t change significantly(Pa0.05).Conclusions RNAi technology targeting hTERT by vector can inhibit the expression of hTERT gene,protein,and increase the apoptosis in HL-60 cell in vitro.