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《Chinese Journal of Reparative and Reconstructive Surgery》 2015-01
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EFFECT OF WNT6 IN PROLIFERATION,DIFFERENTIATION,AND MIGRATION OF BONE MARROW MESENCHYMAL STEM CELLS

CAO Yujing;LV Cuitian;Department of Traumatic Orthopedics,Henan Province Hospital of Traditional Chinese Medicine;Basic Medical College,Henan University of Traditional Chinese Medicine;  
Objective To explore the potential role of WNT6 in the proliferation,differentiation,and migration of bone marrow mesenchymal stem cells(BMSCs). Methods Mouse BMSCs were cultured to the cell fusion of 30%-50%,and divided into different groups. WNT6 knockdown included 3 experiment groups: cells transfected with WNT6 specific short hairpin RNA(sh RNA)(group A1),cells transfected with control sh RNA group(group B1),and nontransfected cells(group C1). WNT6 over-expression included 3 groups: cells transfected with WNT6 recombinant plasmid(group A2),cells transfected with blank vector(group B2),and non-transfected cells(group C2). After transfection,the stably transfected cells were cultured for 48 hours. Cell morphology was observed under inverted microscope; real-time fluorescent quantitative PCR was used to analyze WNT6 m RNA levels; Western blot was used to detect WNT6 and Ki67 protein expressions; cell proliferation was assayed by MTT method,and cell migration was detected by Transwell assay. After cells were cultured in osteogenic differentiation medium for 12 days,the alkaline phosphatase(ALP) activity and calcium deposits were detected by biochemical determination. Results The inverted microscope observation showed that the cell morphology were similar among groups A1,B1,C1,and A2,B2,C2. The WNT6 m RNA and protein levels,Ki67 protein level,cell proliferation,cell migration,ALP activity,and calcium deposition in group A1 were all significantly lower than those in groups B1 and C1(P0.05),but there was no significant difference between groups B1 and C1(P0.05). On the contrary,the above indexes in group A2 were all significantly higher than those in groups B2 and C2(P0.05),but no significant difference was shown between groups B2 and C2(P0.05). Conclusion WNT6 can promote the proliferation and migration,as well as can enhance osteogenic differentiation ability in mouse BMSCs.
【CateGory Index】: R329.2
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