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Comparative transcriptomic analysis reveals adriamycin-induced apoptosis via p53 signaling pathway in retinal pigment epithelial cells.
- Source :
-
Journal of Zhejiang University. Science. B [J Zhejiang Univ Sci B] 2018 Dec.; Vol. 19 (12), pp. 895-909. - Publication Year :
- 2018
-
Abstract
- Objective: This paper applied a transcriptomic approach to investigate the mechanisms of adriamycin (ADR) in treating proliferative vitreoretinopathy (PVR) using ARPE-19 cells.<br />Methods: The growth inhibitory effects of ADR on ARPE-19 cells were assessed by sulforhodamine B (SRB) assay and propidium iodide (PI) staining using flow cytometry. The differentially expressed genes between ADR-treated ARPE-19 cells and normal ARPE-19 cells and the signaling pathways involved were investigated by microarray analysis. Mitochondrial function was detected by JC-1 staining using flow cytometry and the Bcl-2/Bax protein family. The phosphorylated histone H2AX (γ-H2AX), phosphorylated checkpoint kinase 1 (p-CHK1), and phosphorylated checkpoint kinase 2 (p-CHK2) were assessed to detect DNA damage and repair.<br />Results: ADR could significantly inhibit ARPE-19 cell proliferation and induce caspase-dependent apoptosis in vitro. In total, 4479 differentially expressed genes were found, and gene ontology items and the p53 signaling pathway were enriched. A protein-protein interaction analysis indicated that the TP53 protein molecules regulated by ADR were related to DNA damage and oxidative stress. ADR reduced mitochondrial membrane potential and the Bcl-2/Bax ratio. p53-knockdown restored the activation of c-caspase-3 activity induced by ADR by regulating Bax expression, and it inhibited ADR-induced ARPE-19 cell apoptosis. Finally, the levels of the γ-H2AX, p-CHK1, and p-CHK2 proteins were up-regulated after ADR exposure.<br />Conclusions: The mechanism of ARPE-19 cell death induced by ADR may be caspase-dependent apoptosis, and it may be regulated by the p53-dependent mitochondrial dysfunction, activating the p53 signaling pathway through DNA damage.
- Subjects :
- Caspases metabolism
Cell Proliferation
Cell Survival drug effects
Flow Cytometry
Gene Expression Profiling
Gene Expression Regulation
Humans
Membrane Potential, Mitochondrial
Oligonucleotide Array Sequence Analysis
Oxidative Stress drug effects
Phosphorylation
Propidium chemistry
RNA, Small Interfering metabolism
Rhodamines chemistry
Signal Transduction drug effects
Vitreoretinopathy, Proliferative drug therapy
Apoptosis
Doxorubicin pharmacology
Retinal Pigment Epithelium metabolism
Transcriptome
Tumor Suppressor Protein p53 metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1862-1783
- Volume :
- 19
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Journal of Zhejiang University. Science. B
- Publication Type :
- Academic Journal
- Accession number :
- 30507074
- Full Text :
- https://doi.org/10.1631/jzus.B1800408