论文部分内容阅读
【目的】针对Wnt信号通路抑制因子Dickkopf(DKK)1、骨硬化蛋白Sclerostin(Sost)基因构建过表达重组腺病毒载体,观察其对MG63细胞和相关蛋白的影响。【方法】构建DKK1、Sost过表达重组腺病毒载体,将培养好的MG63细胞分为空白对照组、空载病毒组(NC对照组)、过表达DKK1组、过表达Sost组、过表达DKK1+Sost组5组,根据组别不同分别用上述过表达重组腺病毒感染MG63细胞,观察细胞增殖、碱性磷酸酶(ALP)活性、钙离子浓度和骨调节蛋白表达情况。【结果】与空白对照组比较,过表达DKK1组、过表达Sost组、过表达DKK1+Sost组的细胞光密度值、ALP活性值均显著降低,而钙离子浓度显著升高,以过表达DKK1+Sost组变化最明显(P<0.01)。与空白对照组比较,过表达DKK1组、过表达Sost组、过表达DKK1+Sost组的骨保护蛋白(OPG)、低密度脂蛋白受体相关蛋白(Lrp)5、骨形态发生蛋白(BMP)2、结缔组织生长因子(CTGF)、成纤维细胞生长因子(FGF)2、骨桥蛋白(OPN)表达量均降低,而肿瘤坏死因子(TNF)-α表达量则升高,以过表达DKK1+Sost组变化最明显,组间比较差异均有统计学意义(P<0.05或P<0.01)。【结论】过表达DKK1、Sost可降低MG63细胞增殖和ALP活性,升高钙离子浓度,对骨调节蛋白有一定影响,尤以两者同时过表达时的作用最强。
【Objective】 To investigate the effect of Dickkopf (DKK) 1, an inhibitor of Wnt signaling pathway, on the expression of scostostin (Sost) gene in MG63 cells and related proteins. 【Methods】 DKK1 and Sost overexpression recombinant adenovirus vectors were constructed. The cultured MG63 cells were divided into blank control group, NC group, NC group, overexpression DKK1 group, overexpression Sost group and overexpression of DKK1 + Sost group were divided into 5 groups. MG63 cells were infected with the recombinant adenovirus over-expressing respectively according to the groups. The cell proliferation, alkaline phosphatase (ALP) activity, calcium concentration and the expression of osteomodulin were observed. 【Results】 Compared with the blank control group, the ODK and ALP activity of DKK1 overexpression group, Sost overexpression group and DKK1 + Sost overexpression group were significantly lower than those of the blank control group, while the calcium concentration was significantly increased. Over-expression of DKK1 + Sost group the most obvious change (P <0.01). Compared with the blank control group, the expression of osteoprotegerin (OPG), low density lipoprotein receptor associated protein (Lrp) 5, and bone morphogenetic protein (BMP) in overexpression DKK1 group, overexpression Sost group and overexpression DKK1 + The expression of connective tissue growth factor (CTGF), fibroblast growth factor (FGF) 2 and osteopontin (OPN) were decreased, while the expression of tumor necrosis factor (TNF) -α was increased. + Sost group was the most obvious change, the difference between the two groups was statistically significant (P <0.05 or P <0.01). 【Conclusion】 Overexpression of DKK1 and Sost can reduce the proliferation and ALP activity of MG63 cells and increase the concentration of calcium ions, which may affect the expression of osteomodulin.