摘要
本研究旨在探讨低温(28℃)胁迫下JUN(transcription factor AP-1)对HeLa细胞存活率的影响。以体外培养的HeLa细胞为模型,RT-qPCR检测28℃处理1、3、5、10 d时JUN mRNA表达量,Western blot分析JUN的蛋白表达和磷酸化水平,CCK-8法检测细胞存活率。随后通过敲降JUN和过表达JUN、JUN4A,研究不同温度下(37℃、28℃)JUN表达和磷酸化对细胞存活的影响。结果显示,低温处理10 d会导致HeLa细胞中JUN的表达和磷酸化水平增高,Caspase-3的活化增强,细胞存活率逐渐下降。常规温度(37℃)下,JUN过表达和磷酸化对细胞存活是必需的,但是在低温胁迫下敲降JUN,降低JUN磷酸化水平更有利于HeLa细胞的增殖,表明JUN在常温和低温下对HeLa细胞存活发挥不同的影响。本研究为进一步探索哺乳动物细胞在低温胁迫下的调节机制提供部分实验依据。
The aim of this study was to investigate the effect of JUN(transcription factor AP-1)on survival rate of HeLa cells under low temperature(28℃).HeLa cells were exposed to low temperature(28℃)for 1,3,5 and10 days,the JUN mRNA level was detected by RT-qPCR,the protein and phosphorylation levels of JUN were analyzed by Western blot,and cell survival rate was detected by CCK-8 method.To investigate the effects of JUN expression and phosphorylation on cell survival at different temperatures(37℃and 28℃),we knocked down JUN and overexpressed JUN and JUN4A in HeLa cells.The results showed that low temperature stress for 10days increased the expression and phosphorylation levels of JUN in HeLa cells,the activation of Caspase-3 increased,and the cell survival rate decreased gradually.Overexpression and phosphorylation of JUN was required for cell survival under conventional culture conditions(37℃),while knockdown of JUN and reduction of JUN phosphorylation level were more conducive to the proliferation of HeLa cells under low temperature pressure.These results indicated that JUN had different effects on the survival of HeLa cells under normal temperature and low temperature.This study provides some experimental basis for further exploring the regulatory mechanism of mammalian cells under low temperature stress.
作者
张众
李梦佳
韩兵社
张俊芳
Zhang Zhong;Li Mengjia;Han Bingshe;Zhang Junfang(Key Laboratory of Ministry of Education for Exploration and Utilization of Aquatic Genetic Resources,National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai,201306)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2021年第11期3387-3395,共9页
Genomics and Applied Biology
基金
国家自然科学基金项目(81770165)资助
