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LINC00899 promotes osteogenic differentiation by targeting miR-374a and RUNX2 expression.

Experimental and therapeutic medicine (2021-08-28)
Xiaoya Gao, Yun Xue, Kechun Yang
ABSTRACT

Accumulating researches indicate that long non-coding RNAs (lncRNAs) participate in human bone mesenchymal stem cells (hBMSCs) osteogenic differentiation. The present study aimed to investigate the underlying molecular mechanisms of long intergenic non-protein coding RNA 899 (LINC00899) in osteoporosis. Therefore, reverse transcription-quantitative PCR was performed to evaluate the expression levels of LINC00899, microRNA (miR)-374a and runt-related transcription factor 2 (RUNX2) in clinical tissues and hBMSCs. The potential interaction between miR-374a and LINC00899 or RUNX2 was predicted utilizing the StarBase software and verified by luciferase reporter and RNA binding protein immunoprecipitation assays. In addition, alkaline phosphatase activity and Alizarin Red S staining were used to evaluate the osteogenic potential of hBMSCs. The results showed that the expression levels of LINC00899 were gradually increased, whilst those of miR-374a were decreased as the osteogenic induction process progresses. Additionally, the expression of LINC00899 was downregulated in the bone tissues of patients with osteoporosis, where LINC00899 knockdown reduced the expression levels of osteogenesis-related genes osteocalcin (OCN), osteopontin (OPN) and RUNX2 in hBMSCs. LINC00899 was also found to directly target miR-374a, thereby inhibiting its expression. Finally, it was predicted that RUNX2 could be directly targeted by miR-374a, such that miR-374a silencing partially abolished the inhibitory effect of LINC00899 knockdown on the expression of RUNX2, OPN and OCN. Overall, findings of the present study suggested that LINC00899 could facilitate the osteogenic differentiation of hBMSCs and prevent osteoporosis by sponging miR-374a to enhance the expression of RUNX2, which provide a potentially useful therapeutic strategy for patients with osteoporosis.

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Proteinase K from Tritirachium album, lyophilized powder, BioUltra, ≥30 units/mg protein, for molecular biology