Effects of long non-coding RNA H19 on apoptosis of osteoblasts induced by dexamethasone

doi:10.12307/2023.500

Abstract

Abstract: BACKGROUND: The pathogenesis of steroid-induced osteonecrosis of femoral head is still unclear, which is closely related to osteoblast apoptosis.
OBJECTIVE: To investigate the role of long non-coding RNA H19 (lncRNA H19) in dexamethasone-induced osteoblast apoptosis.
METHODS: Mouse osteoblasts (MC3T3-E1) were cultured and divided into control group (no treatment) and dexamethasone group (treated with 1 μmol/L dexamethasone for 24 hours). Western blot was used to detect the protein expression levels of Bax, Bcl-2, and p-ERK1/2/ERK1/2. Flow cytometry and cell viability staining were used to detected cell apoptosis. qRT-PCR was performed to detect the expression of lncRNA H19. lncRNA H19 negative control plasmid (ad-NC) and lncRNA H19 overexpression plasmid (ad-lncRNA H19) were transfected into MC3T3-E1 cells, followed by 24 hours of dexamethasone intervention. Cell apoptosis of MC3T3-E1 was detected using cell viability staining, western blot assay, and flow cytometry. MC3T3-E1 cells were treated with MAPK-ERK pathway inhibitor (PD98059) for 24 hours and western blot was used to detect the protein expression levels of Bax and Bcl-2.
RESULTS AND CONCLUSION: Compared with the control group, the apoptotic rate of MC3T3-E1 cells was increased in the dexamethasone group (P < 0.01), while the expression of lncRNA H19 was decreased (P < 0.01) and the MAPK/ERK signaling pathway was inhibited. Up-regulation of lncRNA H19 in MC3T3-E1 cells reduced the apoptosis of cells (P < 0.01) and activated the MAPK/ERK signaling pathway. MC3T3-E1 cells treated with dexamethasone+ad-lncRNA H19+PD98059 showed an increase in the expression of Bax (P < 0.01) and a decrease in the expression of Bcl-2 (P < 0.01) compared with the cells treated with dexamethasone+ad-lncRNA H19. These findings indicate that overexpression of lncRNA H19 can inhibit dexamethasone-induced apoptosis in MC3T3-E1 cells by activating the MAPK-ERK signaling pathway.

Key words: lncRNA H19, MAPK/ERK signaling pathway, dexamethasone, MC3T3-E1 cell, apoptosis

CLC Number:

Yang Huixia, Bai Zhigang, Chi Hongyang, Ma Tianlong, Ma Shengchao, Jiang Yideng. Effects of long non-coding RNA H19 on apoptosis of osteoblasts induced by dexamethasone[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(28): 4513-4518.

Figures/Tables 4

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