细胞核受体RXRα在2,2',4,4'-四溴二苯醚神经毒性中的作用及其机制

doi:10.3969/j.issn.1004-616x.2020.05.002

癌变·畸变·突变 ›› 2020, Vol. 32 ›› Issue (5): 336-343,349.doi: 10.3969/j.issn.1004-616x.2020.05.002

细胞核受体RXRα在2,2',4,4'-四溴二苯醚神经毒性中的作用及其机制

许迎剑^1,2, 张航², 张建清²

1. 山西医科大学公共卫生学院劳动卫生学系, 山西太原 030001;
2. 深圳市疾病预防控制中心, 广东深圳 518055

收稿日期:2020-02-09 修回日期:2020-08-23 出版日期:2020-10-01 发布日期:2020-10-12
通讯作者: 张建清,E-mail:jianqingzh@szcdc.net E-mail:jianqingzh@szcdc.net
作者简介:许迎剑,E-mail:1305544767@qq.com。
基金资助:
国家自然科学基金（21677103）；深圳市医学重点学科建设经费（SZXK067）

Effect of BDE-47 on nuclear receptor RXRα and toxicity in human neuroblastoma SK-N-SH cells

XU Yingjian^1,2, ZHANG Hang², ZHANG Jianqing²

1. Department of Labour Health, School of Public Health, Shanxi Medical University, Taiyuan 030001, Shanxi;
2. Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, Guangdong, China

Received:2020-02-09 Revised:2020-08-23 Online:2020-10-01 Published:2020-10-12

摘要/Abstract

摘要： 目的：探讨视黄醛X受体（RXRs）为核心的多个核受体在2，2'，4，4'-四溴二苯醚（BDE-47）致神经细胞毒性中的相互作用，揭示受体介导的多溴二苯醚（PBDEs）神经毒性效应的分子机制。方法：构建人神经母细胞瘤细胞SK-N-SH的RXRα基因敲除细胞株及高表达细胞株（分别命名为RXR/KO-SK-N-SH和RXR/OE-SK-N-SH细胞）；CCK-8法分别检测SK-N-SH（野生型）、RXR/KO-SK-N-SH和RXR/OE-SK-N-SH细胞在BDE-47浓度分别为1、5、10、20、50、100、150、200 μmol/L处理12、24、48、72 h时的细胞增殖率；qPCR和Western blot法分别检测上述3种细胞中RXRα、TRα、TRβ、PPARα、PPARγ的mRNA和蛋白表达水平。结果：在染毒时间分别为12、24、48、72 h时，不同浓度BDE-47作用下3种细胞的增殖率比较，差异有统计学意义（P < 0.05）。BDE-47对RXR/KO-SK-N-SH细胞的IC₅₀是野生型和RXR/OE-SK-N-SH细胞的1/2。BDE-47处理3种细胞后，RXRα的mRNA和蛋白表达水平在野生型和RXR/OE-SK-N-SH细胞中显著升高，SK-N-SH细胞和RXR/OE-SK-N-SH细胞中TRα和PPARα的表达升高，此两种受体在RXR/KO-SK-N-SH细胞中表达降低。在本实验所采用的BDE-47处理浓度下，TRβ和PPARγ表达在3种细胞中均无明显变化。结论：RXRα在提高SK-N-SH细胞生存能力中发挥重要作用。RXRα可以介导TRα和PPARα的表达，而BDE-47并未激活或者抑制SK-N-SH细胞TRβ和PPARγ的表达。说明BDE-47对于SK-N-SH细胞的毒性主要是通过激活RXRα，进一步诱导TRα和PPARα的表达而介导的。

关键词: 2,2',4,4'-四溴二苯醚, 人神经母细胞瘤细胞, 视黄醛X受体, 甲状腺激素受体, 过氧化物酶体增殖物激活受体

Abstract: OBJECTIVE: To explore interactions between nucleus receptor RXRs and other nuclear receptors in BDE-47-induced cytotoxicity in human neuroblastoma SK-N-SH cells. METHODS: RXRα-knockout cell lines and RXRα high-expression cell lines from human neuroblastoma SK-N-SH cells (named RXR/KO-SK-N-SH and RXR/OE-SK-N-SH cells,respectively) were constructed. The three cell lines were exposed to different concentrations of BDE-47 (1,5,10,20,50,100,150,200 mol/L) for 12,24,48,72 h,and their proliferation rates were determined by the CCK-8 method. mRNA and protein levels of RXR,TR,TR,PPAR and PPAR were measured by qPCR and Western blot,respectively. RESULTS: Proliferation rates of three cell lines which were exposed to BDE-47 for 12,24,48,72 h were significantly different from each other,(P < 0.05). The IC₅₀ of BDE-47 to RXR/KO-SK-N-SH was 1/2 to SK-N-SH and RXR/OE-SK-N-SH. After treatment with BDE-47,the wild-type and RXR/OE-SK-N-SH cell lines showed significant increase of RXRα mRNA and protein expression levels. Their protein expression levels of TRα and PPARα showed the same changes. However,the knockout SK-N-SH cells showed lower expression level in RXRα. The expression levels of TRβ and PPARγ were not dramatically changed in all three SK-N-SH cell lines after the BDE-47 treatment. CONCLUSION: RXRα played a critical role in improving cell viability. After treatment with BDE-47,RXRα mediated the expression of TRα and PPARα in the three SK-N-SH cell lines,but not the expression of TRβ and PPARγ. These data demonstrate that toxicity of BDE-47 on SK-N-SH cells was medicated by activation of RXRα which enhanced TRα and PPARα expression.

Key words: 2,2',4,4'-tetrabromo diphenyl ether, human neuroblastoma cells, retinoid X receptor, thyroid hormone receptors, peroxisome proliferator-activated receptors

中图分类号:

R994.6

许迎剑, 张航, 张建清. 细胞核受体RXRα在2,2',4,4'-四溴二苯醚神经毒性中的作用及其机制[J]. 癌变·畸变·突变, 2020, 32(5): 336-343,349.

XU Yingjian, ZHANG Hang, ZHANG Jianqing. Effect of BDE-47 on nuclear receptor RXRα and toxicity in human neuroblastoma SK-N-SH cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(5): 336-343,349.

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细胞核受体RXRα在2,2',4,4'-四溴二苯醚神经毒性中的作用及其机制

Effect of BDE-47 on nuclear receptor RXRα and toxicity in human neuroblastoma SK-N-SH cells

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