SHP1 gene induces apoptosis and erythroid differentiation in human erythromyeloblastoid leukemia cell line K562 / 第二军医大学学报

ABSTRACT

Objective: To investigate the role of SHP1 gene in inducing apoptosis and erythroid differentiation in human erythromyeloblastoid leukemia cell line K562. Methods: The full length cDNA of SHP1 gene was cloned by RT-PCR and was subcloned into mammalian expression vector pcDNA3.0. The cDNA sequence of the cloned gene was validated by enzyme digestion and DNA sequencing. Then the recombinant plasmid was used to transfect K562 cells via lipofectin. The apoptosis of K562 cells was examined by Hoechst 33258 staining assay and Annexin V/PI double-labeled assay; the differentiation of K562 cells was observed by benzidine staining and expression of glycophorin A (GPA). Results: RT-PCR and Western blotting analysis showed expression of SHP1 in K562 cells after transfection with pcDAN3-SHP1 plasmid. Apoptotic cells were detected in the K562 cells 48 h after treatment with pcDNA3-SHP1, with the apoptosis rate being 16.84%, which was significantly higher than that in cells transfected with pcDNA3.0 (6.23% , P=0.000). The positive rate of benzidine staining was 14.67% and the positive rate of GPA expression was 19.38% in cells treated with pcNDA3-SHP1, both were significantly different from those in the cells transfected with pcDNA3.0 (P=0.005). Conclusion: Over-expression of SHP1 can effectively induce apoptosis and erythroid differentiation in K562 cells.