Silencing of estrogen receptor beta gene influences the expressions of transforming growth factor beta1 and bone morphogenetic protein 2 in human osteoblasts

doi:10.3969/j.issn.2095-4344.2016.29.001

Abstract

Abstract:

BACKGROUND: There are few studies concerning estrogen receptor β gene, and its mechanism of regulating the bone metabolism is still unclear now.
OBJECTIVE: To analyze the effect of estrogen receptor β (ER β) silencing on the expressions of transforming growth factor β1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) in human osteoblasts
METHODS: There were three groups: blank control group (hFOB 1.19 uninfected with any retrovirus); negative control group (containing invalid interference fragment ER β-shRNA-nc); optimal RNAi group (ER β-shRNA-3). ER β-shRNA retroviral vectors in the optimal RNAi group were used to transfect human osteoblasts followed by resistance screening and cell expansion. MTT assay was used to detect the proliferative activity of ER β-silenced osteoblasts. Then under estrogen intervention, the stable inhibition rate of ER β was determined using western blot assay, and the expressions of TGF-β1 and BMP-2 in human osteoblasts after ER β silencing were detected by RT-PCR technology and western blot assay.
RESULTS AND CONCLUSION: Human osteoblasts that were stably transfected by ER β-shRNA-3 retroviral vector was selected successfully, and ER β silencing had no significant influence on the cell proliferation (P > 0.05). Under the interference of estrogen, the silencing efficiency of ER β protein was (93.11±0.57)% (P < 0.05), and after ER β silencing, the expressions of TGF-β1 and BMP-2 were increased by (26.65±3.81)% and (16.62±1.71)% at mRNA level, and increased by (23.79±3.76)% and (18.08±3.20)% at protein level (both P < 0.05). In conclusion, ER β may play an important role in bone metabolism by regulating the expressions of TGF-β1 and BMP-2.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Estrogen receptor beta, Osteoblasts, Transforming Growth Factor beta 1, Bone Morphogenetic Protein 2

CLC Number:

R318

Deng Ang, Zhang Hong-qi, Guo Chao-feng, Wang Yu-xiang, Gao Qi-le, Tang Ming-xing, Liu Shao-hua, Liu Jin-yang. Silencing of estrogen receptor beta gene influences the expressions of transforming growth factor beta1 and bone morphogenetic protein 2 in human osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(29): 4261-4268.

Figures/Tables 3

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