SPARC Gene Inhibits The Proliferation Of Myelodysplastic Syndrome (MDS) Cell Effect On Proliferation

Objective： To construct a recombinant lentiviral vector carryingSPARC gene, investigate the alteration on the proliferation of MDS cellline SKM-1cells.Methods： SPARC was obtained using SPARC-RNAi-LV,pcDNA-SPARC by PCR. The SPARC gene was cloned into a lentiviralvector to construct a recombinant lentiviral vector carrying SPARC genenamed SPARC-RNAi-LV or pGC-GV-SPARC, which was confirmed byDNA sequencing. SPARC-RNAi-LV and pGC-GV-SPARC was used totransfect human MDS cell line SKM-1.The transfection efficiency, SPARCmRNA and SPARC protein expression were detected by flow cytometry,RT-PCR and Western-blot respectively. The MTS method was used todetermine effects of cell proliferation and low doses Ara-C inhibition ratesof different groups.Results： The recombinant lentiviral vector SPARC-RNAi-LV orpGC-GV-SPARC was confirmed to be correctly by DNA sequencing. Thetransfection efficiency was (64.25±1.42)%, and stable expression. PT-PCR and Western-blot showed that the expression of SPARC was decreased orincreased. MTS results showed that the proliferation of low doses Ara-C onthe inhibition rate of transfection group was significantly higher thannegative group and SKM-1group.Conclusion： Construction of lentiviral vector carrying SPARCsuccessfully, and the expression of SPARC can effectively change the cellproliferation, and overexpress SPARC which combining with low doseAra-C can inhibit the proliferation of SKM-1cell and promote itsapoptosis.