雌激素受体β基因沉默对人成骨细胞转化生长因子β1和骨形态发生蛋白2表达的影响 |
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作者姓名: | 邓 盎 张宏其 郭超峰 王昱翔 高琪乐 唐明星 刘少华 刘金洋 |
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作者单位: | 中南大学湘雅医院脊柱外科,湘雅脊柱外科中心,湖南省长沙市 410008 |
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基金项目: | 国家自然科学基金项目(81271940,81472145) |
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摘 要: | 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
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关 键 词: | 组织构建 骨细胞 雌激素受体β 人成骨细胞 RNA干扰 转化生长因子β1 骨形态发生蛋白2 基因沉默 骨代谢 雌激素 反转录病毒 细胞模型 国家自然科学基金 |
收稿时间: | 2016-04-24 |
Silencing of estrogen receptor beta gene influences the expressions of transforming growth factor beta1 and bone morphogenetic protein 2 in human osteoblasts |
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Authors: | Deng Ang Zhang Hong-qi Guo Chao-feng Wang Yu-xiang Gao Qi-le Tang Ming-xing Liu Shao-hua Liu Jin-yang |
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Institution: | Department of Spinal Surgery, the Affiliated Xiangya Hospital of Central South University, Xiangya Spinal Surgery Center, Changsha 410008, Hunan Province, China |
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Abstract: | |
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Keywords: | Estrogen receptor beta Osteoblasts Transforming Growth Factor beta 1 Bone Morphogenetic Protein 2 |
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