天津医药 ›› 2016, Vol. 44 ›› Issue (12): 1414-1417.doi: 10.11958/20161061

• 细胞与分子生物学 • 上一篇    下一篇

OSAS 模式IH 对肺癌细胞系YAP 以及 P-YAP 表达的影响

薛震1 , 李莲1 , 任方元1 , 姜芳馨2 , 陈宝元1 , 曹洁1△   

  1. 1 天津医科大学总医院呼吸科 (邮编 300052); 2 南开大学生命科学院
  • 收稿日期:2016-09-27 修回日期:2016-11-02 出版日期:2016-12-15 发布日期:2017-01-26
  • 通讯作者: 曹洁 △通讯作者 E-mail: tjcaojie@sina.com E-mail:tjcaojie@sina.com
  • 作者简介: 薛震 (1988), 男, 硕士在读, 主要从事阻塞性睡眠呼吸暂停综合征与肿瘤相关研究
  • 基金资助:
    国家自然科学基金资助项目 (81500070, 81670084); 天津医科大学总医院青年孵育基金项目 (ZYYFY2014011)

Effects of OSAS model IH on the expressions of YAP and P-YAP in lung cancer cell lines

XUE Zhen1 , LI Lian1 , REN Fangyuan1 , JIANG Fangxin2 , CHEN Baoyuan1 , CAO Jie1△   

  1. 1 Department of Respiratory Medicine, Tianjin Medical University General Hospital, Tianjin 300052, China; 2 College of Life Sciences, Nankai University
  • Received:2016-09-27 Revised:2016-11-02 Published:2016-12-15 Online:2017-01-26
  • Contact: CAO Jie △Corresponding Author E-mail: tjcaojie@sina.com E-mail:tjcaojie@sina.com

摘要: 摘要: 目的 探讨阻塞性睡眠呼吸暂停综合征 (OSAS) 模式间歇低氧 (IH) 对肺癌细胞系 A549 细胞 YES 相关蛋白 (YAP) 及磷酸化 YAP (P-YAP) 表达的影响。方法 IH 分别处理 A549 细胞 1 h、 3 h、 6 h (IH1 组、 IH3 组、 IH6 组), IH 频率为: 5% O2 300 s, 21% O2 300 s, 并设置常氧对照组 (N 组)。qRT-PCR 检测 YAP mRNA 表达; Western blot 检测 YAP 及 P-YAP 蛋白表达。结果 IH1 组 (2.50±0.18)、 IH3 组 (4.07±0.25)、 IH6 组 (9.18±0.58) YAP mRNA 的相对表达量显著高于 N 组 (1.00±0.01), 差异有统计学意义 (均 P<0.05), IH1 组、 IH3 组、 IH6 组 YAP 蛋白的表达量较 N 组上调, 并随暴露时间增加而上调; IH1 组、 IH3 组、 IH6 组 P-YAP 蛋白的表达量较 N 组下调, 并随暴露时间增加而下调。结论 YAP 信号通路可能在OSAS 模式IH 促进肿瘤发生发展的过程中起重要的调控作用。

关键词: 睡眠呼吸暂停, 阻塞性, 细胞低氧, 肺癌细胞系 A549 细胞, YES 相关蛋白, 磷酸化 YAP, 实时荧光定量聚合酶链式反应, 蛋白质免疫印迹法

Abstract: Abstract:Objective To explore the effect of obstructive sleep apnea syndrome (OSAS), OSAS- like intermittent hypoxia (IH)on the expression levels of P -YAP and YAP in A549 lung cancer cell lines. Methods A549 cells were treated with IH exposure ( exposed to 5% O2 for 300 seconds and 21% O2 for 300 seconds) for 1, 3 and 6 h (IH1, IH3, IH6) or normoxia exposure (N group). Quantificational real-time PCR was used to measure the mRNA expression levels of YAP. Western blot assay was used to detect the protein expression levels of YAP and P-YAP. Results The mRNA expression levels of YAP were significantly increased with the increase of IH exposure time points in IH1 (2.50±0.18), IH3 (4.07±0.25) and IH6 (9.18±0.58) groups than those in N group (1.00±0.01) (all P<0.05). The protein expression levels of YAP were significantly increased with the increase of IH exposure time points in IH1, IH3 and IH6 groups than those in N group. The protein expression levels of P-YAP were significantly decreased with the increase of IH exposure time points in IH1, IH3 and IH6 groups than those in N group. Conclusion YAP cell signaling plays an important role in the process of OSAS-like IH induced tumor development.

Key words: sleep apnea, obstructive, cell hypoxia, A549 cell, YAP, P-YAP, qRT-PCR, Western blot