癌变·畸变·突变2018，Vol.30Issue(3)：188-193,199,7.DOI:10.3969/j.issn.1004-616x.2018.03.005
矮壮素对小鼠前成骨细胞系MC3T3-E1骨骼发育相关基因及蛋白的影响及其机制
Effects of chlorocholine chloride on expression of skeletal development-related genes and proteins in MC3T3-E1 cells
摘要
Abstract
OBJECTIVE:To investigate the effects of chlorocholine chloride (CCC) on expression of skeletal development-related genes and functional proteins in mouse pre-osteoblast cell line MC3T3-E1.METHODS:MC3T3-E1 cells in logarithmic growth phase were treated with 0,8,40,200,1 000 μg/mL CCC.At different time points (24,48,72 h) after treatment,cytotoxicity was determined by using the MTT assay.At 48 h after treatment,mRNA expression of skeletal and osteogenic development-related genes was detected using qPCR:alkaline phosphatase (ALP),osteocalcin(OCN),growth factor 1 (IGF-1),growth hormone receptor (GHR),RANKL,osteoprotegerin (OPG),macrophage colony-stimulating factor (M-CSF).In addition,expression of functional proteins related to bone development and MAPK signal pathways was determined by Western blot:ALP,GHR,bone morphogenetic protein 2 (BMP2),transcription factor runt-related protein 2 (Runx2).RESULTS:CCC had no significant effect on cytotoxicity of MC3T3-E1 at different time points (24,48,72 h).mRNA expression of ALP,OCN and RANKL in the 1 000 μg/mL groups increased significantly (P＜0.05);the ratio of RANKL and OPG in the 1 000 μg/mL group increased significantly (P＜0.05);mRNA expression of GHR,IGF-1,OPG,M-CSF did not change significantly (P＞0.05).Protein expression of GHR and BMP2 in the 1 000 μg/mL group was significantly decreased,while ALP and RUNX2 did not change significantly.Phosphorylated ERK protein expression decreased and phosphorylated JNK protein expression increased.CONCLUSION:CCC has almost no cytotoxicity on pre-cultured mouse MC3T3-E1 cells,but inhibited the differentiation of MC3T3-E 1 cells into mature osteoblasts and promoted osteoclast differentiation.Moreover,involvement of mitochondria on differentiation of pre-mouse osteoblasts might be related to the MAPK pathway.The decrease of phosphorylated ERK and the increase of phosphorylated JNK might have inhibited migration of MC3T3-E1 to mature osteoblasts differentiation of cells,hence their differentiation to become mature osteoblasts.关键词
矮壮素/成骨细胞/骨骼发育/MAPK通路Key words
chlorocholine chloride/osteoblast/bone development/MAPK pathway分类
医药卫生引用本文复制引用
贾丽霞,张琪,侯晓红,孟庆贺,黄尧,周文娟,郝卫东..矮壮素对小鼠前成骨细胞系MC3T3-E1骨骼发育相关基因及蛋白的影响及其机制[J].癌变·畸变·突变,2018,30(3):188-193,199,7.基金项目
国家自然科学基金(81473002,81773446) （81473002,81773446）
