Overexpression of long non-coding RNA Gm16104 affects osteogenic differentiation of C3H10T1/2 mesenchymal stem cells

doi:10.12307/2024.710

Abstract

Abstract: BACKGROUND: The osteogenic differentiation of mesenchymal stem cells is regulated by a variety of molecules. Long non-coding RNA (lncRNA) has attracted much attention because they can participate in regulating a variety of biological processes, but the regulatory role of lncRNA on osteogenic differentiation of mesenchymal stem cells has not been fully explored.
OBJECTIVE: To explore the effect of lncRNA Gm16104 on osteogenic differentiation of C3H10T1/2 mesenchymal stem cells.
METHODS: The C3H10T1/2 mesenchymal stem cells were induced into osteogenic differentiation by bone morphogenetic protein-2. Alkaline phosphatase staining was performed to identify the osteogenic differentiation of the cells 5 days after osteogenic induction. Expression levels of alkaline phosphatase and lncRNA Gm16104 were detected by qRT-PCR after 0, 1, 3, and 5 days of osteogenic differentiation. After transfection of the overexpression plasmid of pcDNA-Gm16104, the osteogenic differentiation was identified by alkaline phosphatase staining and qRT-PCR 4 days after osteogenic induction. The expression levels of osteogenesis-related signalling pathway proteins were detected by western blot assay.
RESULTS AND CONCLUSION: (1) After 5 days of osteogenic induction, alkaline phosphatase activity was significantly increased. (2) Compared with 0 days, expression levels of the osteogenic marker gene alkaline phosphatase increased and expression levels of Gm16104 decreased after 1, 3, and 5 days of osteogenic induction. (3) Transfection of C3H10T1/2 cells with pcDNA-Gm16104 plasmid significantly increased the expression level of Gm16104. (4) Overexpression of Gm16104 inhibited alkaline phosphatase activity, the expression levels of the osteogenic marker gene alkaline phosphatase and the osteogenesis-related transcription factor Osterix. (5) Overexpression of Gm16104 inhibited phosphorylated protein expression of PI3K and Akt. (6) The above results suggest that overexpression of Gm16104 may inhibit osteogenic differentiation of C3H10T1/2 mesenchymal stem cells through the PI3K/Akt signaling pathway.

Key words: long non-coding RNA, mesenchymal stem cell, Gm16104, bone morphogenetic protein-2, osteogenic differentiation, PI3K, Akt

CLC Number:

Lin Zhanying, Lin Ziyun, Huang Liuyan, Zhang Wenxi, Zuo Changqing. Overexpression of long non-coding RNA Gm16104 affects osteogenic differentiation of C3H10T1/2 mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(31): 4964-4969.

Figures/Tables 6

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