1,25(OH)2D3对人骨肉瘤MG63细胞基质GLA蛋白及Wnt信号通路的影响

摘要

Objective To observe the effects of 1,25-dihydroxyvitamin D3 on Wnt/β-catenin signaling pathways and on the expression of MGP in human osteosarcoma cells line (MG63) in vitro, and to investigate the potential role of vitamin D in postmenopausal osteoporosis.Methods Human osteosarcoma MG63 cells were treated with the following drug intervention respectively:10-8 mol/l 1,25 (OH) 2D3, 200 ng/ml DKK-1, or 10-8 mol/1 1,25 (OH) 2D3 + 200 ng/ml DKK-1 for 48 hours, then total RNA and protein were extracted and examined by Real-Time quantitative RT-PCR and Western blot to measure the mRNA and protein expression of β-catenin, Runx2, LRP5, MGP in MG63 cells.Results 10-8 mol/L 1,25 (OH)2D3 significantly upregulated the expression of classic Wnt/β-catenin signaling pathway associated proteins such as β-catenin, Runx2, LRP5 and MGP.The mRNA expression increased by 2.73-fold, 3.72-fold, 1.53-fold and 2.31-fold, respectively, compared with the control group,and the protein expression increased by 1.13-fold, 1.17-fold, 1.14-fold and 1.21-fold, respectively, compared with the control group, and the differences were statistically significant (P ＜ 0.05).However, DKK1 down-regulated the expression of β-catenin,LRP5, Runx2 and MGP.The mRNA expression decreased by 0.34-fold, 0.52-fold, 0.42-fold and 0.78-fold, respectively, and the protein expression decreased by 0.93-fold, 0.92-fold, 0.87-fold and 0.86-fold, respectively, compared with the control group and the differences were statistically significant (P ＜ 0.05).Conclusion The classic Wnt/β-catenin signaling pathway may play an important part in postmenopausal osteoporosis.1,25 (OH)2D3 may regulated the expression of MGP through the classic Wnt/β-catenin signaling pathway.%目的 观察1,25(OH)2D3对人成骨肉瘤MG63细胞株Wnt信号通路及MGP表达的影响,探讨其在骨质疏松发病中可能的新机制.方法 分别使用10-8 mol/L 1,25(OH)2D3、200 ng/ml Wnt信号通路阻断剂DKK-1、10-8 mol/L 1,25(OH)2D3+200 ng/ml DKK-1干预人骨肉瘤细胞MG63细胞株48 h,使用实时荧光定量RT-PCR及western-blot技术测定Wnt/β-catenin信号通路中相关因子β-catenin、Runx2、LRP5及MGP基因及蛋白表达的影响.结果 (1)10-8 mol/L 1,25(OH)2D3上调MG63细胞中Wnt/β-catenin信号通路中相关因子β-catenin、Runx2、LRP5及MGP的基因表达及蛋白的表达,其中上调基因分别是对照组的2.73倍、3.72倍、1.53倍、2.31倍,差异有统计学意义(P＜0.05);上调蛋白表达分别是对照组的1.13倍、1.17倍、1.14倍、1.21倍,差异有统计学意义(P＜0.05).(2)DKK1下调β-catenin、LRP5、Runx2、MGP的基因及蛋白表达,其中下调基因表达分别是对照组的0.34倍、0.52倍、0.42倍、0.78倍,差异有统计学意义(P＜0.05).下调蛋白表达分别是对照组的0.93倍、0.92倍、0.87倍、0.86倍,差异均有统计学意义(P＜0.05).结论 1,25(OH)2D3有可能通过Wnt/β-catenin信号通路影响MGP的表达.