中国组织工程研究Issue(41)：7188-7198,11.DOI:10.3969/j.issn.2095-4344.2013.41.002
雌激素受体β基因沉默对成骨样MG63细胞骨保护素和RANKL表达的影响
Silenced estrogen receptor beta affects the expressions of osteoprotegerin and receptor activator of nuclear factor-kappa B ligand in osteoblastic MG63 cells
摘要
Abstract
BACKGROUND:Studies concerning how estrogen receptorβparticipates in bone metabolism are few now. OBJECTIVE:To investigate the effect of estrogen receptorβon the expression of osteoprotegerin and receptor activator of nuclear factor-κB ligand in human osteblast-like cells. METHODS:The retrovirus with the most effective interference sequence and non-specific short hairpin RNA was used to transfect human osteoblast-like cellMG63 in order to screen out the stable colon, and then amplified and cultured. The blank control and non-specific short hairpin RNA were used as control, and the stable inhibition rate of estrogen receptorβwas detected. The 17β-estradiol was added into the cells in three groups, that were MG63 cells, short hairpin RNA retrovirus estrogen receptorβ-mediated MG63 cells and negative control short hairpin RNA retrovirus-medicated MG63 cells, in order to detect the expressions of osteoprotegerin and receptor activator of nuclear factor-κB ligand mRNA in human osteoblast-like cells. RESULTS AND CONCLUSION: The human osteoblast-like MG63 cellline was further stably transfected with pRNAT-H1.4/Retro-estrogen receptorβshort hairpin RNA3, and then compared with the blank control and negative control, and found that estrogen receptorβcould express the stable inhibited human osteoblast-like cellline. The inhibition rate of estrogen receptorβmRNA was (88.17±1.17)%(P<0.05), and the inhibition rate of estrogen receptorβprotein was (89.01±1.22)%(P<0.05), indicating that estrogen receptorβgene knockdown human osteoblast-like cellmodels were constructed successful y. After estrogen intervention for 48 hours, the inhibition of MG63 cells with estrogen receptorβcould up-regulate the osteoprotegerin mRNA and protein expression in the blank control group and the negative control group (P<0.05), down-regulate the receptor activator of nuclear factor-κB ligand mRNA and protein expression (P<0.05), and up-regulate the osteoprotegerin receptor activator of nuclear factor-κB ligand expression. The results indicate that estrogen receptorβmay play an important role in bone metabolism through regulating osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio.关键词
组织构建/骨组织构建/雌激素受体β/RNA干扰/人成骨样细胞/反转录病毒载体/骨保护素/RANKL/国家自然科学基金Key words
estrogen receptor beta/osteoprotegrin/receptor activator of nuclear factor kappa B/osteoblasts分类
医药卫生引用本文复制引用
王昱翔,刘金洋,吴建煌,张宏其,郭超峰,唐明星,刘少华,邓盎,高琪乐,邓展生,陈静..雌激素受体β基因沉默对成骨样MG63细胞骨保护素和RANKL表达的影响[J].中国组织工程研究,2013,(41):7188-7198,11.基金项目
国家自然科学基金面上项目(81271940)；湖南省自然科学基金项目(08JJ3057)；湖南省科技厅科技计划一般项目(08FJ3171)；中南大学自由探索计划(2012QNZT122)；湖南省自然科学基金重点项目(12JJ2043) （81271940）
