The role of miR-711 in cardiac cells in response to oxidative stress and its biogenesis: a study on H9C2 cells
Autor: | Bowen Wang, Xiufen Zheng, Tiffany Ni, Fengjun Ling, Tina Mele, Adam Greasley, Ishita Topiwala, Hao Zheng, Cuilin Zhu, Kexiang Liu, Duo Zhao, Qinfeng Zhou |
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Rok vydání: | 2020 |
Předmět: |
Ang-1
0301 basic medicine Cacna1c Antimycin A Apoptosis Mitochondrion medicine.disease_cause Biochemistry 0302 clinical medicine Gene expression Myocytes Cardiac RNA Small Interfering NFКB Membrane Potential Mitochondrial Gene knockdown medicine.diagnostic_test lcsh:Cytology Chemistry NF-kappa B Cobalt Transfection Cell Hypoxia Mitochondria Up-Regulation Cell biology Gene Knockdown Techniques 030220 oncology & carcinogenesis Signal Transduction Programmed cell death Calcium Channels L-Type Cell Survival HIF-1α miR-711 Cell Line Flow cytometry 03 medical and health sciences Angiopoietin-1 medicine Animals lcsh:QH573-671 Molecular Biology Research Hydrogen Peroxide Cell Biology Hypoxia-Inducible Factor 1 alpha Subunit Rats Fibroblast Growth Factors MicroRNAs Oxidative Stress 030104 developmental biology FGF14 Oxidative stress |
Zdroj: | Cellular & Molecular Biology Letters Cellular & Molecular Biology Letters, Vol 25, Iss 1, Pp 1-20 (2020) |
ISSN: | 1689-1392 1425-8153 |
DOI: | 10.1186/s11658-020-00206-z |
Popis: | Background Oxidative stress results in cell apoptosis/death and plays a detrimental role in disease development and progression. Stressors alter the miRNA expression profile and miRNAs play a role in the cell response to stress. We previously showed that miR-711 is significantly over-expressed in extended cold ischemia reperfusion injured hearts in heart transplant. In this study, we aimed to investigate the role of miR-711 in cardiac cell damage in response to oxidative stress and how miR-711 is regulated. Methods Rat cardiac cell line H9c2 cells were cultured and exposed to oxidative conditions (Antimycin A (AA), H2O2, CoCl2, or cold hypoxia/reoxygenation (H/R)) in vitro. H9c2 cells were transfected with miR-711 mimics, miR-711 inhibitors, or small interference RNA, using transfection reagents. The expression of miR-711 was measured by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Cell apoptosis/death was detected by flow cytometry and an IncuCyte system. Mitochondrial damage was detected by measuring the mitochondria membrane potential by flow cytometry. Gene expression was detected by qRT-PCR at the mRNA level and Western blotting and immunocytochemistry staining at the protein level. Results We found that miR-711 was significantly up-regulated in cells treated with H2O2, AA, CoCl2, and cold H/R. Over-expression of miR-711 increased cell apoptosis/death induced by AA and H/R whereas cell death was reduced by miR-711 inhibitors. MiR-711 induced cell death through negative regulation of angiopoietin 1 (Ang-1), fibroblast growth factor 14 (FGF14) and calcium voltage-gated channel subunit alpha1C (Cacna1c) genes. Both knockdown of hypoxia inducible factor 1α (HIF-1α) and inactivation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NFКB) pathway inhibited over-expression of miR-711. Conclusion Oxidative stress increases the expression of miR-711. Over-expression of miR-711 induces cell apoptosis/death. HIF-1α and NFКB regulate miR-711 in H9c2 cells during oxidative stress. miR-711 is a new target for preventing oxidative stress. |
Databáze: | OpenAIRE |
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