Transcriptomic analysis and biological evaluation reveals that LMO3 regulates the osteogenic differentiation of human adipose derived stem cells via PI3K/Akt signaling pathway
Transcriptomic analysis and biological evaluation reveals that LMO3 regulates the osteogenic differentiation of human adipose derived stem cells via PI3K/Akt signaling pathway
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DOI：
10.1007/s10735-021-10047-5
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发表时间：
2022-02-14
影响因子：
3.2
通讯作者：
Pei, Wenye
中科院分区：
文献类型：
--
作者：
Kang, Yue;Pei, Wenye
自体骨移植是治疗骨缺损的常用方法，需要大量的骨细胞。为了开发新的骨组织再生治疗方法，本研究旨在鉴定人脂肪干细胞（hADSCs）成骨的关键基因并探讨其作用机制。下载GSE 63754、GSE 89330和GSE 72429以进行GO功能和KEGG途径分析，构建竞争性内源RNA（ceRNA）网络，构建PPI网络并鉴定枢纽基因。采用定量逆转录聚合酶链反应（RT-PCR）和Western blot检测LMO 3在hADSCs成骨过程中的表达水平。慢病毒转染用于敲低或过表达LMO 3，这使得我们能够研究LMO 3对hADSCs成骨分化的影响。采用Wortmannin检测LMO 3/PI 3 K/Akt轴对hADSCs成骨分化的调控作用。采用裸鼠异位成骨实验研究LMO 3对体内成骨的影响。在本研究中，我们发现LMO 3的表达在hADSCs的成骨分化过程中显著上调。LMO 3基因敲减可显著抑制hADSCs的成骨分化，而LMO 3基因过表达可促进hADSCs的成骨分化。此外，我们发现LMO 3过表达对成骨分化的促进作用与PI 3 K/Akt信号通路的激活有关。用渥曼青霉素抑制PI 3 K/Akt信号通路有效地阻断了LMO 3过表达诱导的成骨分化刺激。总之，基于转录组学分析，我们确定了参与调控hADSC成骨分化的关键基因。此外，我们发现LMO 3可能通过介导PI 3 K/Akt信号通路，对hADSC的成骨分化起正向调节作用。调控LMO 3及其相关通路的表达可能有助于骨再生和组织工程的进展。
Autologous bone transplantation which is a common treatment method for bone defects needs a large quantity of bone cells. In order to develop new treatments to regenerating bone tissues, this research aimed at identifying the key genes and finding their mechanism in human adipose-derived stem cells (hADSCs) osteogenesis. GSE63754, GSE89330 and GSE72429 were downloaded to perform GO functional and KEGG pathway analyses, construct a competing endogenous RNA (ceRNA) network, construct a PPI network and identify hub genes. The expression level of LMO3 during the osteogenesis of hADSCs was examined by quantitative reverse transcription polymerase chain reaction and western blot. Lentivirus transfection was used to knock down or overexpress LMO3, which enabled us to investigate the effect of LMO3 on osteogenic differentiation of hADSCs. Wortmannin were used to identify the mechanism of the LMO3/PI3K/Akt axis in regulating osteogenic differentiation of hADSCs. Moreover, ectopic bone formation in nude mice was used to investigate the effect of LMO3 on osteogenesis in vivo. In this study, we found the expression of LMO3 was significantly upregulated during the osteogenic differentiation of hADSCs. LMO3 knockdown remarkably suppressed osteogenic differentiation of hADSCs, while LMO3 overexpression promoted osteogenic differentiation of hADSCs both in vitro and in vivo. Moreover, we discovered that the enhancing effect of LMO3 overexpression on osteogenic differentiation was related to the activation of PI3K/Akt signaling pathway. Inhibition of PI3K/Akt signaling pathway with wortmannin effectively blocked the stimulation of osteogenic differentiation induced by LMO3 overexpression. In conclusion, based on transcriptomic analysis, we identified key genes involved in regulating the osteogenic differentiation of hADSCs. In addition, we found that LMO3 might act as a positive modulator of hADSC osteogenic differentiation by mediating PI3K/Akt signaling pathway. Manipulating the expression of LMO3 and its associated pathways might contribute to advances in bone regeneration and tissue engineering.