Aim: The molecular mechanism of differentiation in bone marrow mesenchymal stem cells (BMSCs) preserves to be further elucidated. LncRNA HOTTIP has been proven to accelerate osteogenic differentiation, but the regulation mechanism is still unclear.
Methods: The human BMSCs (hBMSCs) were isolated and identified by the antigen CD29, CD34, CD44, CD45, and CD90 through flow cytometry. The osteogenic state was determined by the ALP Detection Kit and Alizarin red staining. The tube formation was observed under a microscope. HOTTIP expression level, DLX2 and TAF15, Wnt/β-catenin pathway, and transcriptional markers in osteogenesis and angiogenesis were examined with Western blot and RT-qPCR, respectively. The combination of TAF15 with lncRNA HOTTIP and DLX2 was detected by RNA immunoprecipitation (RIP) and RNA pulldown assays.
Results: The outcomes revealed that HOTTIP was noticeably up-regulated accompanied by the osteogenic transcriptional factor in the process of osteoblast differentiation and angiogenesis. Besides, HOTTIP enhanced alkaline phosphatase (ALP) activity, accelerated osteogenic differentiation and angiogenesis along with up-regulation of osteogenic and angiogenic-related gene expression, by interaction with TAF15 to stabilize DLX2.
Conclusion: Taken together, our outcomes reveal that lncRNA HOTTIP accelerated osteogenic differentiation and angiogenesis by interaction with TAF15 to stabilize DLX2.
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