Long noncoding RNA LINC00314 facilitates osteogenic differentiation of adipose-derived stem cells through the hsa-miR-129-5p/GRM5 axis via the Wnt signaling pathway

Abstract

Background: Many studies have shown that long noncoding RNAs (lncRNAs) are closely related to the stimulation of osteogenic differentiation of adipose-derived stem cells (ADSCs) and the prevention of osteoporosis. Current research aimed to investigate the novel lncRNA and explored the function and molecular mechanism of the LINC00314/miR-129-5p/GRM5 axis in regulating osteogenic differentiation of ADSCs. Methods: LncRNA and miRNA sequencing was performed in normal and osteogenic differentiation-induced ADSCs (osteogenic group). Abnormally expressed lncRNAs and miRNAs were obtained by the R software and the relative expression of LINC00314, miR-129-5p, and GRM5 during osteogenic induction was measured by RT-PCR. ADSCs were then transfected with pcDNA3.1-sh-LINC00314 and agomiR-129-5p. Alizarin red staining (ARS) and alkaline phosphatase (ALP) staining were performed to identify the mechanism of the LINC00314/miR-129-5p/GRM5 axis in regulating osteogenic differentiation of ADSCs. Results: LINC00314 was significantly upregulated in the group of osteogenic-induced ADSCs. LINC00314 and GRM5 mimics increased the early and late markers of osteogenic differentiation, which manifest in not only the markedly increased ALP activity but also higher calcium deposition, while miR-129-5p mimic had the opposite effects. LINC00314 directly targeted miR-129-5p through luciferase reporter assay, and miR-129-5p suppressed GRM5 expression. Moreover, the LINC00314/miR-129-5p/GRM5 regulatory axis activated the Wnt/β-catenin signaling pathway. Conclusions: LINC00314 confers contributory function in the osteogenic differentiation of ADSCs and thus the LINC00314/miR-129-5p/GRM5 axis may be a novel mechanism for osteogenic-related disease.

miR-129-5p Inhibits Bone Formation Through TCF4

“…MicroRNA inhibited the translation of target gene by binding to its 3′ UTR region ( Lai, 2002 ). In Shi et al’s (2020) study, hsa-miR-129-5p inhibited Wnt/β-catenin pathway via targeting Grm5 . In Li et al (2013) ; Zhang et al (2017) , and Cao et al (2018) , hsa-miR-129-5p inhibited Wnt/β-catenin pathway via targeting Apc , Sox4 , and Wnt5a , respectively.…”

“…miR-129-5p is a miRNA that has been reported as a regulator of cancer development and Alzheimer disease ( Li G. et al, 2019 ; Zeng et al, 2019 ). Shi et al found hsa-miR-129-5p inhibited osteogenic differentiation of adipose-derived stem cells ( Shi et al, 2020 ), while its function on bone formation is still unexplored. To further reveal the correlation between miR-129-5p and osteoporosis, in this study, we have stimulated aging-related osteoporosis in male aging mice model and menopause osteoporosis in female aging menopause mice model.…”

“…miR-129-5p has been reported as a regulator of cancer and neural disease (Li G. et al, 2019;Zeng et al, 2019), whereas studies on its function in bone formation are relatively limited. Shi et al (2020) reported that hsa-miR-129-5p inhibited osteogenic differentiation of adipose-derived stem cells via Wnt/β-catenin pathway, which implied that miR-129-5p may also be an inhibitor of bone formation. Therefore, the reduction of miR-129-5p level in bone tissue might be a potential anabolic strategy for ameliorating osteoporosis.…”

miR-1249-5p regulates the osteogenic differentiation of ADSCs by targeting PDX1

“…Studies have confirmed that several miRNAs can influence the osteogenic differentiation of ADSCs, such as miR-145-5p [ 17 ], miR-143-3p [ 18 ], and miR-129-5p [ 5 ]. Wang et al [ 19 ] reported that overexpression of miR-346 could regulate the Wnt/β-catenin signaling pathway by targeting GSK3b to promote the differentiation of BMSCs into osteoblasts.…”

“…ADSCs can play the function of multidirectional differentiation and self-renewal, and can differentiate into osteoblasts, chondrocytes, adipocytes, nerve cells, and myoblasts under different conditions [ 5 , 6 ]. In addition, ADSCs also have the advantages of easy access, simple sampling, minimally invasive, abundant sources, and relatively small immune rejection [ 7 ].…”