Impact of HOXA10 expression on proliferation and apoptosis of K562 cells

Abstract

Objective To investigate the impact of RNA interference silencing HOXA10 gene on proliferation and apoptosis of K562 cell strain of human chronic myeloid leukemia (CML) by construction of eukaryotic expression vector targeting HOXA10.Methods Based on short hairpin RNA(shRNA) oligo that was designed and compounded by the selected specific small interference RNA (siRNA) targeting HOXA 10,the eukaryotic expression vector of pGPHI-GFP-Neo-HOXA10 was successfully constructed and sequenced,and the cationic liposome was then used to transfect K562 cells.There were cell control group (equivalent cells and culture medium only),negative control group (negative control plasmid transfected by liposome)and experimental group (pGPHI- GFP- Neo- HOXA10 transfected by liposome).The HOXA10 mRNA expression was detected by RT-PCR at 24 h after transfection,cell proliferation by MTT at 24,48 and 72 h for cell inhibition ratio,and the apoptosis by flow cytometry at 48 h in all the groups.Results The vector pGPHI-GFP-Neo-HOXA10 was successfully constructed and used to transfect K562 cells.The transfection vector in experimental group could effectively decrease the expression level of HOXA10 mRNA [ (38.86±4.49)％ vs (88.52±9.24)％,(86.75±7.38)％,P＜0.05] as compared with that in cell control and negative control group,with no difference between negative control and cell control group (P＞0.05).Compared with negative control group,the experimental group had a significant decrease in cell proliferation capacity but an increase in cell inhibition ratio at 24,48 and 72 h after transfection [ (39.92±0.74)％ vs (7.98±5.52)％;(55.62±1.18)％ vs (8.27±3.45)％; (66.30±1.26)％ vs (8.63±3.58)％; all P＜0.05].Moreover,the apoptosis rate was significantly increased in experimental group as compared with that in control and negative control groups [ (22.29± 1.67)％ vs (9.82±0.69)％,(10.14±0.96)％,P＜0.05],however,no difference was found between negative control and cell control group (P＞0.05).Conclusion Since eukaryotic expression vector pGPHI-GFP-Neo-HOXA10 can effectively silence the expression of HOXA10 in K562 cells,pGPHI-GFP-Neo-HOXA10 may significantly inhibit proliferation of K562 cell and induce its apoptosis. Key words: RNA,small interfering; K562 cells; Genes,homeobox; Cell proliferation; Apoptosis