Circ_KIAA0922通过调控miR-148a-3p/SMAD5轴,激活TGF-β信号通路,调控Saos-2细胞增殖和成骨分化

IF 1.1 Q2 MEDICINE, GENERAL & INTERNAL
Shanshan Zhang, Yongtao Zhang, Dan Yang, Wei Zhi, Junfeng Li, Meilin Liu, Yanqin Lu, Jinxiang Han
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引用次数: 0

摘要

环状rna (circRNAs)正在成为人类疾病的重要调节因子,但其在骨质疏松症(OP)中的功能尚不清楚。本研究的目的是确定circ_KIAA0922在体外Saos-2细胞成骨分化中的可能分子机制,以及circ_KIAA0922与miR-148a-3p和SMAD家族成员5 (SMAD5)之间的相互作用。Circ_KIAA0922、miR-148a-3p和SMAD5通过瞬时转染过表达。采用双荧光素酶报告系统分析circ_KIAA0922、miR-148a-3p和SMAD5的组合。此外,采用RT-qPCR或western blot分析检测circ_KIAA0922、miR-148a-3p、SMAD5、骨钙素(OCN)和矮子相关转录因子2 (RUNX2)的水平。茜素红染色分析circ_KIAA0922、miR-148a-3p和SMAD5控制下的成骨分化程度。我们发现circ_KIAA0922敲低抑制Saos-2细胞的增殖和成骨分化。Circ_KIAA0922直接靶向miR-148a-3p, miR-148a-3p抑制逆转了Circ_KIAA0922敲低对Saos-2细胞增殖和成骨分化的影响。SMAD5的过表达促进了Saos-2细胞的增殖和成骨分化,减弱了miR-148a-3p对细胞增殖和成骨分化的抑制作用。综上所述,circ_KIAA0922在体外通过circ_KIAA0922/miR-148a-3p/ SMAD5轴促进Saos-2细胞增殖和成骨分化,从而为circ_KIAA0922促进成骨分化的机制提供了新的认识。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Circ_KIAA0922 regulates Saos-2 cell proliferation and osteogenic differentiation by regulating the miR-148a-3p/SMAD5 axis and activating the TGF-β signaling pathway
Circular RNAs (circRNAs) are emerging as important regulators in human disease, but their function in osteoporosis (OP) is not sufficiently known. The aim of this study was to identify the possible molecular mechanism of circ_KIAA0922 in osteogenic differentiation of Saos-2 cells in vitro and the interactions among circ_KIAA0922, miR-148a-3p, and SMAD family member 5 (SMAD5). Circ_KIAA0922, miR-148a-3p, and SMAD5 were overexpressed by transient transfection. Dual-luciferase reporter assay system was used to analyze the combination among circ_KIAA0922, miR-148a-3p, and SMAD5. In addition, the levels of circ_KIAA0922, miR-148a-3p, SMAD5, osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) were detected using RT-qPCR or western blot analysis. Alizarin red staining was performed to analyze the degree of osteogenic differentiation under the control of circ_KIAA0922, miR-148a-3p, and SMAD5. We found that circ_KIAA0922 knockdown inhibited the proliferation and osteogenic differentiation of Saos-2 cells. Circ_KIAA0922 directly targeted miR-148a-3p, and miR-148a-3p inhibition reversed the effects of circ_KIAA0922 knockdown on the proliferation and osteogenic differentiation of Saos-2 cells. Overexpression of SMAD5 promoted the proliferation and osteogenic differentiation of Saos-2 cells and attenuated the inhibitory effect of miR-148a-3p on cell proliferation and osteogenic differentiation. In conclusion, circ_KIAA0922 facilitated Saos-2 cell proliferation and osteogenic differentiation via the circ_KIAA0922/miR-148a-3p/ SMAD5 axes in vitro, thus providing insights into the mechanism of osteogenic differentiation by circ_ KIAA0922.
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来源期刊
Intractable & rare diseases research
Intractable & rare diseases research MEDICINE, GENERAL & INTERNAL-
CiteScore
2.10
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29
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