Overexpression of stromaI cell-derived factor-1 promotes the proIiferation and migration of bone marrowmesenchymaI stem cells in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND: The stromal cell-derived factor-1 (SDF-1)/C-X-C chemokine receptor type 4 (CXCR4) signaling pathway cannot only improve the migration ability of bone marrow mesenchymal stem cells (BMSCs), but also restrain BMSCs apoptosis, increase BMSCs survival and improve the proliferation activity of BMSCs. OBJECTIVE: To construct a rat BMSCs line with SDF-1α overexpression and to explore its influence on the proliferation and migration of BMSCs in vitro. METHODS: The SDF-1α overexpression vector (pNL-SDF-1α-IRES2-EGFP) was constructed. The lentivirus particles were packaged by transferring pNL-SDF-1α-IRES2-EGFP, pNL-IRES2-EGFP and GV-118-SDF-1α-siRNA into 293T cells. The BMSCs lines with SDF-1α overexpression in SDF-1α-BMSCs group, null-BMSCs group and siRNA-BMSCs group were established by transfecting SDF-1α-lentiviru, null-lentivirus and siRNA-lentivirus into BMSCs respectively. The expression of SDF-1α at mRNA and protein levels in BMSCs was evaluated by RT-PCR and western blot assay, respectively. The influence of SDF-1α on proliferation and migration of BMSCs were evaluated by MTT and Transwell migration experiment respectively. RESULTS AND CONCLUSION: The pNL-SDF-1α-IRES2-EGFP recombinant plasmid was successfully constructed, which was proved by sequencing results. EGFP was strongly expressed in 293T cells and BMSCs in all groups after 48 hours in lentivirus transfection. SDF-1α at mRNA and protein levels were highly expressed in the SDF-1α-BMSCs group, but the expression was significantly inhibited in the siRNA-BMSCs group. The proliferative ability of BMSCs was strengthened in the SDF-1α-BMSCs group, and SDF-1α was found to significantly promote the transmembrane migration of BMSCs. The migration index of BMSCs incubated with anti-SDF-1α multi-antibodies was restrained markedly. To conclude, the lentivirus vector cannot only infect BMSCs efficiently but also make SDF-1 expresse stably in BMSCs. The overexpression of SDF-1α can improve the proliferation and migration abilities of BMSCs.