Regulatory effects of LncRNA MEG3 on the proliferation and apoptosis of nucleus pulposus cells in rat intervertebral disc

doi:10.12307/2021.118

Abstract

Abstract:

BACKGROUND: Injury of nucleus pulposus cells in the intervertebral disc is one of the important causes of degenerative disc disease. The mechanism of longchain non-coding RNA (LncRNA) in intervertebral disc degeneration has not been clarified.

OBJECTIVE: To investigate the effect of LncRNA MEG3 on the proliferation and apoptosis of nucleus pulposus cells in rat intervertebral discs and its regulatory effect on phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway.

METHODS: Primary cultured rat nucleus pulposus cells were treated with interleukin 1β (IL-1β) to establish a cell model. There were five groups in this experiment, including control group, IL-1β group, IL-1β+pcDNA group, IL-1β+pcDNA-MEG3 group, and IL-1β+pcDNA-MEG3+LY294002 group. Real-time quantitative PCR was used to detect the expression of MEG3. MTT was used to measure cell proliferation. Flow cytometry was used to detect apoptosis rate.Western blot was used to detect the expression of Ki-67, proliferating cell nuclear antigen (PCNA), Bax, Cleaved Caspase-3, PI3K, p-PI3K, Akt, and p-Akt.
RESULTS AND CONCLUSION: Compared with the control group, in the IL-1β group, the expression of MEG3 in the cells was significantly reduced (P < 0.05), the cell viability was significantly reduced (P < 0.05), the apoptosis rate was significantly increased (P < 0.05), the levels of Ki-67, PCNA, p-PI3K, and p-Akt proteins were significantly reduced (P < 0.05), and the levels of Bax and Cleaved Caspase-3 proteins were significantly increased (P<0.05). Compared with the IL-1β+pcDNA group, in the IL-1β+pcDNA-MEG3 group, the cell viability was significantly increased (P < 0.05), the levels of Ki-67, PCNA, p-PI3K, and p-Akt proteins were significantly increased (P < 0.05), the apoptosis rate was significantly reduced (P < 0.05), and the levels of Bax and Cleaved Caspase-3 proteins were significantly reduced (P < 0.05). Compared with the IL-1β+pcDNA-MEG3 group, in the IL-1β+pcDNA-MEG3+LY294002 group, the cell viability was significantly reduced (P < 0.05), the apoptosis rate was significantly increased (P < 0.05), the levels of Ki-67 and PCNA proteins were significantly reduced (P < 0.05), and the levels of Bax and Cleaved Caspase-3 proteins were significantly increased (P < 0.05). To conclude, overexpression of MEG3 may promote IL-1β-induced proliferation and inhibit apoptosis in rat nucleus pulposus cells by activating the PI3K/Akt signaling pathway.

Key words: LncRNA MEG3, PI3K/Akt signaling pathway, rat, intervertebral disc, nucleus pulposus cells, proliferation, apoptosis

CLC Number:

Cheng Ning, Zhang Zhizhong. Regulatory effects of LncRNA MEG3 on the proliferation and apoptosis of nucleus pulposus cells in rat intervertebral disc[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(26): 4106-4111.

Figures/Tables 7

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