线粒体核糖体蛋白MRPS28参与碳离子辐照诱导A549细胞凋亡研究  

Mitochondrial Ribosomal Protein MRPS28 Participates in Apoptosis of A549 Cells Induced by Carbon Ion Irradiation

作　　者：黄国敏 张金花[1,2,3,4,5] 窦志慧 包兴婷 柳佳娣 徐度玲 李鸿岩 张红[1,2,3,4,5] HUANG Guomin;ZHANG Jinhua;DOU Zhihui;BAO Xingting;LIU Jiadi;XU Duling;LI Hongyan;ZHANG Hong(Department of Medical Physics,Institute of Modern Physics,Chinese Academy of Sciences,Lanzhou 730000,China;Key Laboratory of Heavy Ion Irradiation Biology and Medicine of Chinese Academy of Sciences,Lanzhou 730000,China;Key Laboratory of Basic Research on Heavy Ion Irradiation Application in Medicine,Lanzhou 730000,China;College of Life Sciences,University of Chinese Academy of Sciences,Beijing 101408,China;Advanced Energy Science and Technology Guangdong Laboratory,Huizhou 516029,Guangdong,China)

机构地区：[1]中国科学院近代物理研究所医学物理研究室,兰州730000 [2]中国科学院重离子辐照生物学与医学重点实验室,兰州730000 [3]重离子辐照医学应用基础研究重点实验室,兰州730000 [4]中国科学院大学核科学与技术学院,北京101408 [5]先进能源科学与技术广东省实验室,广东惠州516029

出　　处：《原子核物理评论》2024年第4期1066-1074,共9页Nuclear Physics Review

基　　金：国家重点研发计划资助项目(2018YFE0205100);国家自然科学基金资助项目(11875061,12005042);中央政府引导地方科技发展资金(甘肃省)项目(GSCK2021-51-11);甘肃省自然科学基金项目(20JR5RA550);广东省自然科学基金项目(2021A151010027)。

摘　　要：线粒体核糖体蛋白(Mitochondrial Ribosomal Proteins,MRPs)是一类既受细胞核调控,又受线粒体调控的蛋白质。我们研究发现非小细胞肺癌A549细胞暴露于碳离子辐照后细胞中MRPS28表达量下降。采用siRNA敲低MRPS28并利用CRISPR-Cas9方法构建稳定敲低MRPS28的A549细胞系。结果发现,相对于对照组,MRPS28敲低能够降低A549细胞的增殖及存活率;同时增加细胞中ROS含量,降低线粒体膜电位,并促进细胞凋亡。用Western Blot检测凋亡相关蛋白的表达发现,敲低MRPS28能够通过p53介导的线粒体凋亡途径诱导细胞凋亡。此外,在A549细胞中MRPS28基因表达下调与碳离子辐照的联合作用显著抑制了细胞的增殖并降低了存活率,同时加速了辐照诱导的细胞凋亡进程。这一结果表明,MRPS28表达水平的下调可能增强了A549细胞对碳离子辐照的敏感性。本研究初步证明MRPS28低表达参与碳离子辐照诱导的A549细胞凋亡,并增强了A549细胞对碳离子束的辐射敏感性。Mitochondrial ribosomal proteins(MRPs)are a category of proteins that are regulated by both cell nucleus and mitochondria.In this study,we have observed a down-regulation of the mitochondrial ribosomal protein MRPS28 in non-small cell lung cancer A549 cells following exposure to carbon ion irradiation.Utilizing small interfering RNA(siRNA)-mediated gene silencing to specifically target MRPS28 in A549 cells,and employing the CRISPR/Cas9 system for genomic editing,we have successfully generated a stable A549 cell line with attenuated MRPS28 expression.Comparative analysis with the control group revealed that the down-regulation of MRPS28 expression resulted in a significant reduction in both the proliferative capacity and the survival rate of A549 cells.Simultaneously,the knockdown of MRPS28 leads to an increase in intracellular reactive oxygen species(ROS)content,a decrease in mitochondrial membrane potential,and the promotion of apoptosis.Upon employing Western Blot to evaluate the expression profiles of proteins implicated in apoptosis,it was observed that the down-regulation of MRPS28 induces apoptosis through the activation of the p53-dependent mitochondrial apoptotic cascade.Furthermore,in the A549 cell line,the combined effect of MRPS28 gene expression down-regulation and carbon ion irradiation significantly inhibited cellular proliferative activity and reduced survival capacity,while also accelerating the process of irradiation-induced apoptosis.These results suggest that down-regulation of MRPS28 expression may enhance the radio-sensitivity of A549 cells to carbon ion radiation.The present investigation provides initial evidence that down-regulation of MRPS28 contributes to apoptosis in A549 cells induced by carbon ion irradiation,thereby augmenting the radio-sensitivity of A549 cells to carbon ion.

关 键 词：碳离子辐照 A549细胞 线粒体核糖体蛋白 凋亡 ROS p53 

分 类 号：Q691[生物学—生物物理学]