Abstract: Objective To investigate the effects of SHP-1 gene on the proliferation, apoptosis, cell cycle,and colony formation of K562 cells line in vitro. Methods K562 cells were infected with the lentiviral plasmids carrying the specified retroviral vector(pEX-SHP-1-puro-Lv105) or the control vector(pEX-EGFPpuro-Lv105). The expression of SHP-1 mRNA and protein were determined by SYBR Green-based qRTPCR and Western blot. The proliferation of cells was detected by CCK-8 assay, and the apoptosis rate and cell cycle were assessed by flow cytometry. Besides, cell morphology was observed by light and electron microscopy. Cell apoptosis was detected by TUNEL method. Cell colonies were cultured and the number of colonies was compared between groups. Results CCK-8 assays showed that the cell proliferation rate was sharply reduced in K562SHP-1 cells (0.35±0.02), compared with K562EGFP cells(0.47±0.05) at the third day of culture(P=0.011). Flow cytometry analysis showed that K562SHP-1 cells were arrested in G0/G1 phase(56.37±2.27), compared with K562EGFP cells(31.67±3.21) (P=0.000) at the fifth day of culture; and the percentage of apoptotic K562SHP-1 cells(26.13±1.16) was significantly higher than that of apoptotic K562EGFP cells(9.00 ±1.22)(P=0.000). The colony forming ability of K562SHP-1 cells (26.3±5.2) was decreased compared with K562EGFP cells (54.7±8.6), (P=0.000). SHP-1 overexpression caused a slight decrease of BCR-ABL1 protein in K562SHP-1 cells compared with K562EGFP cells(P=0.040). Conclusion The overexpression of SHP-1 gene could inhibit the proliferation and colony formation of K562 cells, induce cell apoptosis, result in cell cycle arrest at G0/G1 phase, and decrease the level of BCR-ABL1 protein.