Abstract:
Objective To investigate the mechanism of the effect of soybean isoflavone (ISO) on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells by regulating Wnt-1 gene expression.
Methods Primary marrow mesenchymal stem cells were isolated and cultured, and treated with ISO at different concentrations (40, 60, 80 μg/ μL) (ISO-L group, ISO-M group, ISO-H group) . Liposome-based transfection technology was used to transfect pcDNA and pcDNA-Wnt-1 into bone marrow mesenchymal stem cells (pcDNA group and pcDNA-Wnt-1 group) , respectively, and to transfect si-NC and si-Wnt-1 into bone marrow mesenchymal stem cells, followed by the treatment with ISO (ISO+si-NC group and ISO+si-Wnt-1 group) . MTT was used to detect cell proliferation. The effects of ISO on Wnt-1 expression were detected by RT-qPCR and Western blot, respectively. RT-qPCR and Western blot were used to detect the expression levels of ALP, OPN, and Runx2, respectively. The independent sample t test was used for the comparison between two groups, and the one-way analysis of variance and pair LSD-t test were used for the comparison between multiple groups.
Results Compared with the control group, the cell viability of ISO-L group, ISO-M group and ISO-H group was significantly increased [24 h: (0.17±0.02) vs (0.21±0.02) , (0.25±0.02) , (0.29±0.03) , 48 h: (0.26±0.03) vs (0.39±0.03) , (0.48±0.04) , (0.53±0.05) ; 72 h: (0.37±0.03) vs (0.54±0.04) , (0.65±0.06) , (0.78±0.07) ], mRNA and protein levels of ALP, OPN, Runx2 and Wnt-1 in bone marrow mesenchymal stem cells were also significantly increased [ALP mRNA: (1.00±0.06) vs (1.74±0.16) , (2.53±0.21) , (3.06±0.29) ; ALP protein: (0.41±0.04) vs (0.62±0.06) , (0.74±0.05) , (0.83±0.07) . OPN mRNA: (1.01±0.08) vs (1.52±0.15) , (2.11±0.19) , (2.74±0.25) ; OPN protein: (0.34±0.03) vs (0.51±0.05) , (0.63±0.06) , (0.76±0.07) . Runx2 mRNA: (1.01±0.08) vs (1.52±0.15) , (2.11±0.19) , (2.74±0.25) ; Runx2 protein: (0.34±0.03) vs (0.51±0.05) , (0.63±0.06) , (0.76±0.07) ], and they all showed a gradually increasing trend (P < 0.05) . Compared with the pcDNA group, the cell viability of the pcDNA-Wnt-1 group [24 h: (0.18±0.02) vs (0.24±0.02) ; 48h: (0.25±0.03) vs (0.51±0.05) ; 72h: (0.36±0.03) vs (0.72±0.06) ], ALP mRNA [ (1.01±0.08) vs (2.89±0.27) ] and protein [ (0.40±0.04) vs (0.79±0.07) ], OPN mRNA [ (0.99±0.07) vs (2.53±0.25) ] and protein [ (0.33±0.03) to (0.71± 0.06) ], Runx2 mRNA [ (1.00±0.06) vs (2.14±0.21) ] and protein [ (0.21±0.02) vs (0.62±0.06) ] expression levels all showed a gradual increase trend, and the difference was statistically significant (all P < 0.05) . Compared with the ISO+si-NC group, the cell viability of the ISO+si-Wnt-1 group [24 h: (0.25±0.03) vs (0.19±0.02) ; 48 h: (0.56±0.05) vs (0.31±0.03) ; 72 h: (0.82±0.08) vs (0.49±0.04) ], ALP mRNA [ (3.14±0.31) vs (1.67±0.16) ] and protein [ (0.87±0.08) vs (0.51±0.05) ], OPN mRNA [ (2.79±0.26) vs (1.43±0.13) ]and protein [ ( 0.79±0.08) vs (0.43±0.04) ], Runx2 mRNA [ (2.36±0.24) vs (1.35±0.13) ]and protein [ (0.68±0.07) vs (0.34±0.03) ] expression levels all showed a gradual decrease trend, and the differences were statistically significant (all P < 0.05) .
Conclusion ISO can enhance the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells by regulating the expression of Wnt-1.
Key words:
Soy bean isoflavone,
Wnt-1,
Bone marrow mesenchymal stem cells,
Proliferation,
Osteogenic differentiation
Yidong Liu, Yufeng Jia, Yan Wang, Yang Liu, Jinyue Ning. Upregulation of Wnt-1 gene expression by soybean isoflavones to promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2021, 11(05): 292-297.