Comparative transcriptomic analysis reveals adriamycin-induced apoptosis via p53 signaling pathway in retinal pigment epithelial cells

作     者：Yu-chen LIN Ze-ren SHEN Xiao-hui SONG Xin LIU Ke YAO 

作者机构：Eye Centerthe Second Affiliated HospitalZhejiang University School of MedicineHangzhou 310009China Key Laboratory of Ophthalmology of Zhejiang ProvinceHangzhou 310009China 

出 版 物：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 (浙江大学学报（英文版）B辑（生物医学与生物技术）)

年 卷 期：2018年第19卷第12期

页      面：895-909页

核心收录：

学科分类：1002[医学-临床医学] 100212[医学-眼科学] 10[医学] 

基　　金：Project supported by the Zhejiang Province Key Research and Development Program(No.2015C03042) China 

主　　题：Adriamycin Proliferative vitreoretinopathy Retinal pigment epithelial p53 Apoptosis 

摘      要：Objective: This paper applied a transcriptomic approach to investigate the mechanisms of adriamycin(ADR) in treating proliferative vitreoretinopathy(PVR) using ARPE-19 cells. 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 H2 AX(γ-H2 AX), phosphorylated checkpoint kinase 1(p-CHK1), and phosphorylated checkpoint kinase 2(p-CHK2) were assessed to detect DNA damage and repair. Results: ADR could significantly inhibit ARPE-19 cell proliferation and induce caspasedependent 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-H2 AX, p-CHK1, and p-CHK2 proteins were up-regulated after ADR exposure. 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.