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.

自体骨移植是治疗骨缺损的常用方法，需要大量的骨细胞。为了开发再生骨组织的新疗法，本研究旨在识别关键基因并发现它们在人类脂肪干细胞 (hADSCs) 成骨中的作用机制。下载 GSE63754、GSE89330 和 GSE72429 以执行 GO 功能和 KEGG 通路分析，构建竞争性内源性 RNA (ceRNA) 网络，构建 PPI 网络并识别中心基因。通过定量逆转录聚合酶链反应和蛋白质印迹检测hADSCs成骨过程中LMO3的表达水平。慢病毒转染用于敲低或过表达 LMO3，这使我们能够研究 LMO3 对 hADSCs 成骨分化的影响。Wortmannin 用于鉴定 LMO3/PI3K/Akt 轴在调节 hADSCs 成骨分化中的机制。此外，裸鼠异位骨形成被用来研究LMO3对体内成骨的影响。在本研究中，我们发现在 hADSCs 的成骨分化过程中 LMO3 的表达显着上调。LMO3 敲低显着抑制了 hADSCs 的成骨分化，而 LMO3 过表达促进了体外和体内 hADSCs 的成骨分化。此外，我们发现LMO3过表达对成骨分化的增强作用与PI3K/Akt信号通路的激活有关。渥曼青霉素抑制 PI3K/Akt 信号通路有效阻断了 LMO3 过表达诱导的成骨分化的刺激。总之，基于转录组学分析，我们确定了参与调节 hADSCs 成骨分化的关键基因。此外，我们发现 LMO3 可能通过介导 PI3K/Akt 信号通路作为 hADSC 成骨分化的正调节剂。操纵 LMO3 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。