Col24α1基因低表达对成骨细胞碱性磷酸酶及矿化作用的影响

摘要

目的:探讨细胞外基质基因Col24α1在MC3T3-E1细胞成骨样分化过程中的表达情况,及其对分化过程中碱性磷酸酶活性和细胞矿化作用的影响.方法:培养MC3T3-E1细胞并诱导成骨分化,从基因及蛋白水平观察Col24α1在成骨细胞分化过程中的表达情况;构建Col24α1低表达慢病毒载体包装,滴度测定,细胞转染效率的验证;分别转染MC3T3-E1细胞,与空白组对照观察Col24α1沉默后碱性磷酸酶活性变化,细胞成骨矿化的影响.结果:Col24α1在成骨细胞在分化过程中表达随着矿化过程的进展而增加.使用慢病毒介导的RNA干扰(RNAi)技术,可以在MC3T3-E1成骨细胞中成功降低Col24α1表达,发现沉默Col24α1表达显著降低ALP活性和细胞矿化作用.结论:Col24α1可以调节成骨细胞的ALP活性和细胞矿化作用,其异常表达可能与骨骼病变相关.%Objective:To observe expression of the extracellular matrix gene,Col24α1 in osteoblast-like MC3T3 E1 cells during the differentiation,alkaline phosphatase activity and cell mineralization.Method:Cultured MC3T3-E1 cells and induced osteogenic differentiation,observed Col24α1 expression in osteoblast differentiation process at gene and protein levels; packaged Col24α1 low expressing vector,titer check,cell transfection efficiency verification; MC3T3-E1 cells were transfected with lentiviral,observed Col24α1 silence and alkaline phosphatase activity on cells and bone mineralization.Col24α1 in osteoblasts expressed during differentiation with the increase in the mineralization process.Lentivirus-mediated RNA interference (RNAi) technology,the expression of Col24α1 in MC3T3-El osteoblasts was successfully reduced,ALP activity and cell mineralization were reduced by silence Col24α1 expression significantly.Conclusion:Col24α1 can adjust osteoblast ALP activity and cell mineralization,its abnormal expression may be associated with bone lesions.