Effect of imatinib combined with P27 gene clone on chronic myeloid leukemia cell line K562

Objective To explore the effect of imatinib combined with P27 gene clone on the proliferation,cycle and apoptosis of chronic myeloid leukemia cell line K562. Methods P27 gene was amplified from peripheral blood mononuclear cells by RT-PCR,and its sequences were confirmed. P27-pcDNA3.1 vector was constructed and then transfected into P27 gene-deleted K562 cell line by lipofectine. After screened with G418,P27-pcDNA3.1-K562 cell clone that stably expressed P27 was isolated. P27 protein was identified by Western blot. The cell survival rate was tested by MTT,cell cycle and apoptosis were tested by flow cytometry. Results The expression of P27 protein could be detected by Western blot in P27-pcDN3.1-K562 cells. A strong inhibition of cell proliferation was observed in P27-pcDNA3.1-K562 cell compared with the control K562 cell. P27-pcDNA3.1-K562 cells increased apparently in G0/G1 phase but declined greatly in S phase. Cell cycle was arrested in G0/G1 phase. The percentage of apoptosis greatly increased after P27-pcDNA3.1-K562 cells were combined with imatinib and the cell survival rate greatly declined. Conclusion Imatinib combined with P27 gene clone has a synergetic effect on inhibiting cell proliferation and promoting apoptosis of K562 cell.