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CLC number: R3

On-line Access: 2018-12-03

Received: 2018-07-30

Revision Accepted: 2018-09-28

Crosschecked: 2018-11-16

Cited: 0

Clicked: 2787

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Yu-chen Lin

https://orcid.org/0000-0002-2679-2849

Ke Yao

https://orcid.org/0000-0002-6764-7365

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Journal of Zhejiang University SCIENCE B 2018 Vol.19 No.12 P.895-909

http://doi.org/10.1631/jzus.B1800408


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


Author(s):  Yu-chen Lin, Ze-ren Shen, Xiao-hui Song, Xin Liu, Ke Yao

Affiliation(s):  Eye Center, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China; more

Corresponding email(s):   xlren@zju.edu.cn

Key Words:  Adriamycin, Proliferative vitreoretinopathy, Retinal pigment epithelial, p53, Apoptosis


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Yu-chen Lin, Ze-ren Shen, Xiao-hui Song, Xin Liu, Ke Yao. Comparative transcriptomic analysis reveals adriamycin-induced apoptosis via p53 signaling pathway in retinal pigment epithelial cells[J]. Journal of Zhejiang University Science B, 2018, 19(12): 895-909.

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author="Yu-chen Lin, Ze-ren Shen, Xiao-hui Song, Xin Liu, Ke Yao",
journal="Journal of Zhejiang University Science B",
volume="19",
number="12",
pages="895-909",
year="2018",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1800408"
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%T Comparative transcriptomic analysis reveals adriamycin-induced apoptosis via p53 signaling pathway in retinal pigment epithelial cells
%A Yu-chen Lin
%A Ze-ren Shen
%A Xiao-hui Song
%A Xin Liu
%A Ke Yao
%J Journal of Zhejiang University SCIENCE B
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1800408

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T1 - Comparative transcriptomic analysis reveals adriamycin-induced apoptosis via p53 signaling pathway in retinal pigment epithelial cells
A1 - Yu-chen Lin
A1 - Ze-ren Shen
A1 - Xiao-hui Song
A1 - Xin Liu
A1 - Ke Yao
J0 - Journal of Zhejiang University Science B
VL - 19
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SP - 895
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PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1800408


Abstract: 
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 H2AX (γ-H2AX), 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 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. 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.

比较转录组学分析揭示阿霉素经p53信号通路诱导视网膜色素上皮细胞凋亡

目的:通过比较阿霉素(ADR)作用的视网膜色素上皮细胞(ARPE-19)和正常ARPE-19间的差异表达基因和信号通路,探究ADR治疗玻璃体视网膜增殖性疾病的潜在机制.
创新点:首次运用基因芯片技术通过比较转录组学分析探究ADR促进ARPE-19细胞凋亡的机制.
方法:采用磺酰罗丹明B(sulforhodamine B,SRB)比色法和碘化丙啶(PI)单染结合流式细胞术检测ADR对ARPE-19细胞的增殖抑制作用;通过基因芯片技术筛选ADR作用的ARPE-19 细胞(实验组)和正常ARPE-19细胞(对照组)间的差异表达基因和相关信号通路;用JC-1染色结合流式细胞术和Bcl2/Bax蛋白表达比率检测线粒体功能;通过检测γ-H2AX、p-CHK1、 p-CHK2等蛋白表达量分析DNA的损伤和修复.
结论:ADR通过启动DNA损伤反应,引起p53信号通路依赖的线粒体功能失调并激活caspase依赖的凋亡,最终导致ARPE-19细胞死亡.

关键词:阿霉素;玻璃体视网膜增殖性疾病;视网膜色素上皮层;p53;凋亡

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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