Function And Mechanism Of LncRNA-2271 On Osteogenic Differentiation Of BMSCs

Objective: To study the effect of lnc R-2271 on osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs),and to explore the mechanism of lnc R-2271 on the differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods: BMSCs was isolated and cultured from bone marrow of patients with femoral neck fracture and divided into three groups.The groups of over expression of lnc R-2271,low expression of lnc R-2271,and the negative control group was set up,each group was cultured by osteogenic inducer.After transfection with 48 h,the expression of lnc R-2271 in three groups of cells was detected by real-time quantitative PCR.The activity of alkaline phosphatase(ALP)was detected by enzyme labeled method after transfection for 7d and 14 d.ELISA method was used to detect the content of bone morphogenetic protein(BMP2),western blot method was used to detect the expressions of Smad1/5/8 and Runx2;After transfection with 21 d,BMSCs culture mediums were stained with alizarin red,and the mineralized nodule area ratio was calculated by Image J image processing software.To analyze the mechanism of lnc R-2271 regulate the osteogenic differentiation of BMSCs.Results: compared with the control group,the expression of lnc R-2271 of over-expressed group was significantly higher than that of the control group(P<0.01)after transfection with 48 h,and the content of lnc R-2271 in the low expression group was significantly lower than that in the negative control group(P<0.01).After intervention with 7d and 14 d,the content of BMP2,Smad1/5/8 and Runx2 in lnc R-2271 over-expressed group was significantly higher than that in negative control group and lnc R-2271 low expression group(P<0.01).The content of BMP2,Smad1/5/8 and Runx2 in low expression group was significantly lower than that in negative control group(P<0.01).After intervention with 7d and 14 d,the expression of ALP in lnc R-2271 over expression group was significantly higher than that in the control group and the lnc R-2271 low expressed group(P<0.01),and the activity of ALP in the lnc R-2271 low expressed group was significantly lower than that in the control group(P<0.01);After intervention with 21 d,alizarin red staining showed low expression group were weakly positive,control group were positive,while the over expression group showed strong positive,calcium deposition of over expression group was significantly higher than control group and low expression group(P<0.01),and calcium deposition of low expression group was significantly lower than the control group(P<0.01).Conclusion: 1.lncR-2271 can promote the expression of BMP2,Smad1/5/8 and Runx2 protein in vitro,and ultimately promote the osteogenic differentiation of BMSCs,which is of great significance for bone regeneration and bone formation in BMSCs.2.lnc R-2271 may promote osteogenic differentiation of BMSCs through BMP2-Smad1/5/8 signaling pathway.