Expression and significance of miR - 20b in retinal photoreceptor cells exposed to PCB1254
Expression and significance of miR - 20b in retinal photoreceptor cells exposed to PCB1254
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PCB1254暴露的视网膜感光细胞中miR-20b的表达及意义
DOI：
10.18632/aging.102360
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发表时间：
2019-10
期刊：
影响因子：
--
通讯作者：
Meiling Tong
中科院分区：
文献类型：
--
作者：
Xin Zhang;Qingyu Zhang;Yue Jiang;Shuchun Zhang;Qin Hong;Xirong Guo;Xia Chi;Meiling Tong
以往的研究表明，PCB1254对斑马鱼视网膜发育有不良影响，但其背后的基本机制尚不清楚。本研究的目的是探讨多氯联苯诱导视网膜发育不良的分子机制。采用RT-qPCR、免疫印迹、HE染色和免疫荧光检测mRNA和蛋白水平的表达。在661w细胞中进行CCK-8、caspase-3、流式细胞术等功能实验，并利用斑马鱼模型进一步研究miR - 20b的功能作用。结果显示PCB1254暴露抑制了661w细胞的增殖，增加了661w细胞的凋亡，并发现了视网膜发育相关基因（SWS1、CRX、Rho）、miR-20b表达与PCB1254暴露之间的剂量-反应关系。我们证实miR-20b通过构建双荧光素酶报告基因靶向FGF2和GRB2，抑制细胞功能以及PCB1254。在miR-20b过表达的斑马鱼模型中，我们发现视网膜形态异常，表现为感光细胞稀疏、不规则，感光细胞层较厚。我们的研究结果首次证明多氯联苯通过miR-20b靶向MAPK/ERK信号，影响视网膜细胞发育并导致视力障碍。
Previous studies have shown that PCB1254 has an adverse effect on zebrafish retinal development, but the basic mechanism behind it is not clear. The purpose of this study was to investigate the molecular mechanisms of PCB-induced retinal dysplasia. RT-qPCR, immunoblotting, HE staining and immunofluorescence were adopted to detect the expression at mRNA and protein level. Functional experiments were carried out in 661w cells including CCK-8 assay, caspase-3 assay, and the flow cytometry, while the functional role of miR - 20b was further investigated by using the zebrafish model. The result showed that PCB1254 exposure inhibited cell proliferation and increased the apoptosis of the 661w cells, and the dose–response relationship between the retinal development-related genes (SWS1, CRX, Rho), miR-20b expression and PCB1254 exposure was also discovered. We confirmed that miR-20b targeted FGF2 and GRB2 by constructing a dual luciferase reporter gene and suppressed the cell function as well as PCB1254. In the miR-20b overexpression ebrafish model, we found abnormal retinal morphology characterized by sparse and irregular photoreceptor cells and the thick photoreceptor cell layers. Our results demonstrate for the first time that PCBs target the MAPK/ERK signaling through miR-20b, affecting retinal cell development and leading to visual impairment.
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影响因子：
4.5
作者：
Tran NM;Zhang A;Zhang X;Huecker JB;Hennig AK;Chen S
通讯作者：
Chen S
影响因子：
7.3
作者：
Belov AA;Mohammadi M
通讯作者：
Mohammadi M
DOI：
10.1387/ijdb.150310ht
发表时间：
2015
期刊：
The International journal of developmental biology
影响因子：
--
作者：
Cheng-Yung Lin;Hsing-Yen Huang;Po-nien Lu;Chien-Wei Lin;Kuan-Ming Lu;H. Tsai
通讯作者：
Cheng-Yung Lin;Hsing-Yen Huang;Po-nien Lu;Chien-Wei Lin;Kuan-Ming Lu;H. Tsai
影响因子：
2
作者：
通讯作者：
--
影响因子：
5.8
作者：
Rosalinda Gioia;A. Akindele;S. A. Adebusoye;K. Asante;S. Tanabe;A. Buekens;A. Sasco
通讯作者：
Rosalinda Gioia;A. Akindele;S. A. Adebusoye;K. Asante;S. Tanabe;A. Buekens;A. Sasco