Silencing of estrogen receptor beta gene influences the expressions of transforming growth factor beta1 and bone morphogenetic protein 2 in human osteoblasts / 中国组织工程研究

ABSTRACT

BACKGROUND:There are few studies concerning estrogen receptorβgene, and its mechanism of regulating the bone metabolism is stil unclear now. OBJECTIVE:To analyze the effect of estrogen receptorβ(ERβ) silencing on the expressions of transforming growth factorβ1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) in human osteoblasts METHODS:There were three groupsblank control group (hFOB 1.19 uninfected with any retrovirus);negative control group (containing invalid interference fragment ERβ-shRNA-nc);optimal RNAi group (ERβ-shRNA-3). ERβ-shRNA retroviral vectors in the optimal RNAi group were used to transfect human osteoblasts fol owed by resistance screening and cel expansion. MTT assay was used to detect the proliferative activity of ERβ-silenced osteoblasts. Then under estrogen intervention, the stable inhibition rate of ERβwas determined using western blot assay, and the expressions of TGF-β1 and BMP-2 in human osteoblasts after ERβsilencing were detected by RT-PCR technology and western blot assay. RESULTS AND CONCLUSION:Human osteoblasts that were stably transfected by ERβ-shRNA-3 retroviral vector was selected successful y, and ERβsilencing had no significant influence on the cel proliferation (P>0.05). Under the interference of estrogen, the silencing efficiency of ERβprotein was (93.11±0.57)%(P<0.05), and after ERβsilencing, 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, ERβmay play an important role in bone metabolism by regulating the expressions of TGF-β1 and BMP-2.