Silenced estrogen receptor beta affects the expressions of osteoprotegerin and receptor activator of nuclear factor-kappa B ligand in osteoblastic MG63 cells

doi:10.3969/j.issn.2095-4344.2013.41.002

Abstract

Abstract:

BACKGROUND: Studies concerning how estrogen receptor β participates in bone metabolism are few now.
OBJECTIVE: To investigate the effect of estrogen receptor β on 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 cell MG63 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 receptor β was detected. The 17β-estradiol was added into the cells in three groups, that were MG63 cells, short hairpin RNA retrovirus estrogen receptor β-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 cell line was further stably transfected with pRNAT–H1.4/Retro-estrogen receptor β short hairpin RNA3, and then compared with the blank control and
negative control, and found that estrogen receptor β could express the stable inhibited human osteoblast-like cell line. The inhibition rate of estrogen receptor β mRNA was (88.17±1.17)% (P < 0.05), and the inhibition rate of estrogen receptor β protein was (89.01±1.22)% (P < 0.05), indicating that estrogen receptor β gene knockdown human osteoblast-like cell models were constructed successfully. After estrogen intervention for 48 hours, the inhibition of MG63 cells with estrogen receptor β could 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 receptor β may play an important role in bone metabolism through regulating osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio.

CLC Number:

R318

Wang Yu-xiang, Zhang Hong-qi, Guo Chao-feng, Tang Ming-xing, Liu Shao-hua, Deng Ang, Gao Qi-le, Deng Zhan-sheng, Chen Jing, Liu Jin-yang, Wu Jian-huang. Silenced estrogen receptor beta affects the expressions of osteoprotegerin and receptor activator of nuclear factor-kappa B ligand in osteoblastic MG63 cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(41): 7188-7198.

Figures/Tables 10

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