Transition of WT1 isoforms inhibits proliferation and promotes apoptosis of human leukemia cell line HL-60

AIM: To study the potential effects of exogenous Wilms tumor 1(WT1) isoforms on the proliferation and apoptosis of human leukemia cell line HL-60.METHODS: WT1(17AA-/KTS-) gene obtained by RT-PCR was cloned into a PCDH1-MCS1-EF1-copGFP plasmid.The recombinant plasmid was confirmed by enzyme digestion and sequencing,and was transfected into HL-60 cells by LipofectamineTM 2000.The stable transformants were selected by G418 screening.WT1(17AA-/KTS-) expression was identified by real-time fluorescence quantitative RT-PCR and Western blotting.The proliferation of the cells was measured by MTT assay.The apoptosis of the cells was determined by morphological observation and flow cytometry analysis.RESULTS: The eukaryotic expression vector PCDH1-MCS1-EF1-copGFP-WT1(17AA-/KTS-) was successfully constructed.The recombinant cells exhibited high mRNA and protein levels of WT1(17AA-/KTS-).The growth of recombinant cells was slower than that of HL-60 cells transfected with control vector and normal HL-60 cells.After exposed to As2O3 at 2 μmol/L for 48 h,both recombinant cells and control cells exhibited the morphological characteristics of apoptosis,but the former was more typical than the latter.The apoptosis was enhanced in the recombinant cells after the cells were exposed to As2O3 for 24 h.CONCLUSION: Exogenous WT1(17AA-/KTS-) isoform inhibits the proliferation and promotes the apoptosis of leukemic cells.