Objective      To construct sperm-associated antigen 6 (SPAG6)-short hairpin RNA (shRNA)-expressing lentiviral vector targeting SPAG6 gene

and investigate the effect of SPAG6 gene silencing on the proliferation and apoptosis of myelodysplastic syndrome (MDS) cell line SKM-1.      Methods      Three kinds of shRNA-containing plasmids for interfering with SPAG6 gene expression were designed

constructed

and transfected into 293T cells to obtain lentiviral vector expressing SPAG6-shRNA. Then

the lentiviral vector was transferred into SKM-1 cells

and the transfection efficiency was detected by flow cytometry. The interference efficiency of SPAG6 gene was determined by quantitative reverse transcription PCR and Western blotting

and the most effective shRNA-containing plasmid was screened. Cell proliferation was examined by CCK-8 assay

and cell apoptosis rate was detected by flow cytometry.      Results      SPAG6-shRNA-expressing lentiviral vector was successfully constructed

and the average transfection efficiency was higher than 60%. Quantitative reverse transcription PCR and Western blot results showed that SPAG6-shRNA3-expressing lentiviral vector was the most effective vector for knock-down of SPAG6 gene. After SPAG6 gene interference

the proliferation of SKM-1 cells in the interference group was inhibited significantly

and the inhibitory rate [(60.18±0.37)%] was higher than that in the negative control group (P0.05). Meanwhile

the cell apoptosis rate of the interference group [(16.32±5.80)%] was significantly higher than that of the negative control group [(4.28±0.88)%

P=0.024] and that of the blank control group [(3.08±1.50)%

P=0.019].      Conclusion      SPAG6-shRNA-expressing lentiviral vector can effectively down-regulate the expression of SPAG6 gene

and significantly inhibit cell proliferation and promote apoptosis of SKM-1 cells.