Primary cilium acts as an oxygen sensor in PC12 cells

doi:10.3785/j.issn.1008-9292.2017.12.07

J Zhejiang Univ (Med Sci)

2017, Vol. 46

Issue (6): 618-624 DOI: 10.3785/j.issn.1008-9292.2017.12.07

Primary cilium acts as an oxygen sensor in PC12 cells

GAO Yuhai1(

),QI Xinzhu2,ZHOU Jian1,WANG Xin2,CHEN Keming1,MA Huiping2,*(

)

1. Institute of Orthopaedics, Lanzhou General Hospital of PLA, Lanzhou 730050, China
2. Key Laboratory of Prevention and Treatment for the Plateau Environmental Damage of PLA, Lanzhou General Hospital of PLA, Lanzhou 730050, China

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Abstract

Objective: To investigate the function of primary cilium as an oxygen sensor in PC12 cells. Methods: The PC12 cells were transfected with IFT88 siRNA. The nuclear translocation of hypoxia inducible factor-1α (HIF-1α), nuclear factor erythroid-2 related factor 2 (Nrf2), and ciliogenesis were observed by immunofluorescence staining; and the mRNA expressions of HIF-1α, Nrf2, vascular endothelial growth factor (VEGF) and superoxide dismutase (SOD) were detected by real-time RT-PCR. Results: The ciliogenesis was inhibited in PC12 cells transfected with IFT88 siRNA. In hypoxia group and scramble control group, nuclear translocations of HIF-1α and Nrf2 were observed and mRNA expressions of HIF-1α, Nrf2, VEGF were increased, and those of SOD were decreased. While in PC12 cells transfected with IFT88 siRNA, nuclear translocations of HIF-1α and Nrf2 were not observed, and mRNA expressions of HIF-1α, Nrf2, VEGF were inhibited, and mRNA expression of SOD was increased. Conclusion: Primary cilia may act as an oxygen sensor to transfer the information related to hypoxia and oxidative stress into cells, activating intracellular defense mechanism against the hypoxic injuries.

Key words： PC12 cells Cilia Receptors, sensory Anoxia/physiopathology Mechanotransduction, cellular RNA, small interfering

Received: 05 March 2017 Published: 25 December 2017

CLC:

Q74

Corresponding Authors: MA Huiping E-mail: gyh1389@126.com;mahuipingcxr@aliyun.com

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	GAO Yuhai
	QI Xinzhu
	ZHOU Jian
	WANG Xin
	CHEN Keming
	MA Huiping

Cite this article:

GAO Yuhai,QI Xinzhu,ZHOU Jian,WANG Xin,CHEN Keming,MA Huiping. Primary cilium acts as an oxygen sensor in PC12 cells. J Zhejiang Univ (Med Sci), 2017, 46(6): 618-624.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2017.12.07 OR http://www.zjujournals.com/med/Y2017/V46/I6/618

初级纤毛细胞外氧感受器功能和机制研究

目的: 探索初级纤毛是否为PC12细胞的细胞外氧感受器。方法: 通过小干扰RNA（siRNA）干扰细胞纤毛内转运蛋白88的表达抑制体外培养PC12细胞初级纤毛的发生；免疫荧光染色法观察缺氧诱导因子1α（HIF-1α）、核因子E2相关因子2（Nrf2）核转位和PC12细胞初级纤毛生长情况；实时定量RT-PCR法检测HIF-1α、Nrf2、血管内皮生长因子（VEGF）和超氧化物歧化酶（SOD）基因表达。结果: siRNA干扰组细胞的初级纤毛被抑制。与常氧对照组比较，缺氧对照组和阴性对照组细胞的HIF-1α和Nrf2发生了明显的核转位，细胞内HIF-1α、Nrf2、VEGF基因表达量增加，SOD表达量下降；siRNA干扰组细胞在缺氧状态下HIF-1α和Nrf2未发生核转位，细胞内HIF-1α、Nrf2、VEGF基因表达受到抑制，而SOD表达量升高。结论: PC12细胞的初级纤毛可能具有细胞外氧感受器的作用，可以感知细胞外环境中的缺氧和氧化应激状态，并通过一定途径将缺氧信号传导至细胞内，进而启动细胞抗缺氧机制。

关键词： PC12细胞, 纤毛, 感受器, 感觉, 缺氧/病理生理学, 动力传导, 细胞, RNA, 小分子干扰

Tab 1 Primer sequences for real-time RT-PCR

Fig 1 Morphology changes of PC12 cells after hypoxia

Fig 2 Observation of the results of siRNA interference in PC12 cells at 24 h after transfection

Fig 3 Effects of siRNA interference on primary cilia

Fig 4 HIF-1α and Nrf2 nuclear translocation in PC12 cells after hypoxia culture for 18 hours

Fig 5 mRNA expressions of HIF-1α, Nrf2, VEGF and SOD in PC12 cells


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