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MicroRNA-494 inhibition protects nucleus pulposus cells from TNF-α-induced apoptosis by targeting JunD.
Biochimie. 2015 Aug; 115:1-7.B

Abstract

BACKGROUND

Human nucleus pulposus cell (HNPC) apoptosis plays an important role in the development of intervertebral disc degeneration (IVDD). Our previous research revealed that among all of the dysregulated microRNAs in the degenerated nucleus pulposus tissues of patient with IVDD, miRNA-494 (miR-494) is the most significantly increased. However, the influence of miR-494 HNPC apoptosis has not been confirmed.

OBJECTIVE

This study was designed to evaluate the effect of miR-494 on the HNPC apoptosis induced by TNF-α and to explore the possible mechanism of this process.

METHODS

First, HNPCs were stimulated with TNF-α at different concentrations (0 ng/ml, 10 ng/ml, 50 ng/ml, or 100 ng/ml) for 0 h, 8 h, 16 h, or 24 h. Annexin V-PE/7-AAD assays and real-time quantitative PCR were used to detect the cell apoptosis rates and miR-494 expression. Second, we successfully knocked down endogenous miR-494 in HNPCs via lentiviral antigomiR-494 vector infection and then stimulated with TNF-α (100 ng/ml, 16 h). The rates of apoptosis and miR-494 expression were then detected again. Additionally, a dual-luciferase reporter assay and western blotting were used to determine whether JunD is a target of miR-494. Finally, western blotting was used to analyze the expression of cytochrome C.

RESULTS

We found that the rate of apoptosis increased with concentration, time (p < 0.05) and miR-494 expression (p < 0.05). The rate of apoptosis in the 100 ng/ml, 16 h group appeared to be suitable. After transfection, the apoptosis rate and miR-494 expression were significantly decreased in the antigomiR-494+TNF-α group compared to the controls (p < 0.05). We also revealed that JunD is a target of miR-494. Western blotting analysis demonstrated that treatment with the lentiviral antigomiR-494 vector resulted in increased expression of JunD (p < 0.05) and decreased expression of cytochrome C (p < 0.05).

CONCLUSION

These results indicated that miR-494 is a novel regulator of HNPC apoptosis induced by TNF-α. The knock-out of miR-494 expression protected the HNPCs from apoptosis via the up-regulation of JunD, which was possibly mediated via cytochrome C apoptotic signaling. These findings suggest that the miR-494/JunD signaling pathway might represent a novel therapeutic target for the prevention of IVDD.

Authors+Show Affiliations

Tian Jin Medical University, No.22, Qixiangtai Road, Heping District, Tianjin 300070, China; Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China.Tian Jin Medical University, No.22, Qixiangtai Road, Heping District, Tianjin 300070, China; Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China.Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China. Electronic address: maxinlong86@163.com.Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China.Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China.Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China.Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China.Tian Jin Hospital, No.406, Jiefangnan Road, Hexi District, Tianjin 300211, China.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25906693

Citation

Wang, Tao, et al. "MicroRNA-494 Inhibition Protects Nucleus Pulposus Cells From TNF-α-induced Apoptosis By Targeting JunD." Biochimie, vol. 115, 2015, pp. 1-7.
Wang T, Li P, Ma X, et al. MicroRNA-494 inhibition protects nucleus pulposus cells from TNF-α-induced apoptosis by targeting JunD. Biochimie. 2015;115:1-7.
Wang, T., Li, P., Ma, X., Tian, P., Han, C., Zang, J., Kong, J., & Yan, H. (2015). MicroRNA-494 inhibition protects nucleus pulposus cells from TNF-α-induced apoptosis by targeting JunD. Biochimie, 115, 1-7. https://doi.org/10.1016/j.biochi.2015.04.011
Wang T, et al. MicroRNA-494 Inhibition Protects Nucleus Pulposus Cells From TNF-α-induced Apoptosis By Targeting JunD. Biochimie. 2015;115:1-7. PubMed PMID: 25906693.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - MicroRNA-494 inhibition protects nucleus pulposus cells from TNF-α-induced apoptosis by targeting JunD. AU - Wang,Tao, AU - Li,Pengfei, AU - Ma,Xinlong, AU - Tian,Peng, AU - Han,Chao, AU - Zang,Jiacheng, AU - Kong,Jingbo, AU - Yan,Hongliang, Y1 - 2015/04/20/ PY - 2014/12/06/received PY - 2015/04/11/accepted PY - 2015/4/25/entrez PY - 2015/4/25/pubmed PY - 2016/4/9/medline KW - Apoptosis KW - Intervertebral disc degeneration KW - JunD KW - MicroRNA KW - TNF-α SP - 1 EP - 7 JF - Biochimie JO - Biochimie VL - 115 N2 - BACKGROUND: Human nucleus pulposus cell (HNPC) apoptosis plays an important role in the development of intervertebral disc degeneration (IVDD). Our previous research revealed that among all of the dysregulated microRNAs in the degenerated nucleus pulposus tissues of patient with IVDD, miRNA-494 (miR-494) is the most significantly increased. However, the influence of miR-494 HNPC apoptosis has not been confirmed. OBJECTIVE: This study was designed to evaluate the effect of miR-494 on the HNPC apoptosis induced by TNF-α and to explore the possible mechanism of this process. METHODS: First, HNPCs were stimulated with TNF-α at different concentrations (0 ng/ml, 10 ng/ml, 50 ng/ml, or 100 ng/ml) for 0 h, 8 h, 16 h, or 24 h. Annexin V-PE/7-AAD assays and real-time quantitative PCR were used to detect the cell apoptosis rates and miR-494 expression. Second, we successfully knocked down endogenous miR-494 in HNPCs via lentiviral antigomiR-494 vector infection and then stimulated with TNF-α (100 ng/ml, 16 h). The rates of apoptosis and miR-494 expression were then detected again. Additionally, a dual-luciferase reporter assay and western blotting were used to determine whether JunD is a target of miR-494. Finally, western blotting was used to analyze the expression of cytochrome C. RESULTS: We found that the rate of apoptosis increased with concentration, time (p < 0.05) and miR-494 expression (p < 0.05). The rate of apoptosis in the 100 ng/ml, 16 h group appeared to be suitable. After transfection, the apoptosis rate and miR-494 expression were significantly decreased in the antigomiR-494+TNF-α group compared to the controls (p < 0.05). We also revealed that JunD is a target of miR-494. Western blotting analysis demonstrated that treatment with the lentiviral antigomiR-494 vector resulted in increased expression of JunD (p < 0.05) and decreased expression of cytochrome C (p < 0.05). CONCLUSION: These results indicated that miR-494 is a novel regulator of HNPC apoptosis induced by TNF-α. The knock-out of miR-494 expression protected the HNPCs from apoptosis via the up-regulation of JunD, which was possibly mediated via cytochrome C apoptotic signaling. These findings suggest that the miR-494/JunD signaling pathway might represent a novel therapeutic target for the prevention of IVDD. SN - 1638-6183 UR - https://www.unboundmedicine.com/medline/citation/25906693/MicroRNA_494_inhibition_protects_nucleus_pulposus_cells_from_TNF_α_induced_apoptosis_by_targeting_JunD_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0300-9084(15)00115-7 DB - PRIME DP - Unbound Medicine ER -