Omicron变异株XBB.1.16刺突蛋白对人非小细胞肺癌细胞的生物学功能的影响

R734.2; 目的 探讨新型冠状病毒Omicron变异株XBB.1.16刺突蛋白(XBB.1.16-S)感染人非小细胞肺癌细胞H1299-ACE2的病毒学特征及其对H1299-ACE2细胞线粒体受损的影响.方法 以亲本pCMV3-SARS-CoV-2-S质粒为模板构建XBB.1.16-S基因真核表达载体(pCMV3-XBB.1.16-S质粒)后转染H1299-ACE2细胞,比较两者诱导细胞形成合胞体的能力.利用双质粒系统包装SARS-CoV-2-S和XBB.1.16-S假病毒后分别感染H1299-ACE2细胞,测定感染后宿主细胞荧光素酶强度并采用Western blot实验检测细胞中S蛋白的...

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Published in中国癌症防治杂志 Vol. 16; no. 3; pp. 271 - 276
Main Authors 房廖鑫, 亢小峰, 薛春源, 潘露, 陈佳欣, 汤传昊, 孙丽颖, 徐小洁, 杜祎萌
Format Journal Article
LanguageChinese
Published 100071 北京 中国人民解放军军事科学院军事医学研究院 2024
Subjects
Mitochondrion
H1299-ACE2细胞
Non-small cell lung cancer
XBB.1.16
Omicron
线粒体
非小细胞肺癌
H1299-ACE2 cells
Online AccessGet full text
ISSN1674-5671
DOI10.3969/j.issn.1674-5671.2024.03.02

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Abstract R734.2; 目的 探讨新型冠状病毒Omicron变异株XBB.1.16刺突蛋白(XBB.1.16-S)感染人非小细胞肺癌细胞H1299-ACE2的病毒学特征及其对H1299-ACE2细胞线粒体受损的影响.方法 以亲本pCMV3-SARS-CoV-2-S质粒为模板构建XBB.1.16-S基因真核表达载体(pCMV3-XBB.1.16-S质粒)后转染H1299-ACE2细胞,比较两者诱导细胞形成合胞体的能力.利用双质粒系统包装SARS-CoV-2-S和XBB.1.16-S假病毒后分别感染H1299-ACE2细胞,测定感染后宿主细胞荧光素酶强度并采用Western blot实验检测细胞中S蛋白的切割效率,采用MitoTracker Green探针观察染色细胞的线粒体形态,采用流式细胞仪检测细胞的线粒体膜电位(mitochondrial membrane potential,MMP)水平和活性氧(reactive oxygen species,ROS)水平.结果 与亲本pCMV3-SARS-CoV-2-S质粒相比,pCMV3-XBB.1.16-S质粒的T19I、V83A、G142D等关键氨基酸突变位点突变成功,且转染H1299-ACE2细胞后所形成的合胞体面积明显减小(P＜0.0001).与SARS-CoV-2-S相比,XBB.1.16-S感染H1299-ACE2细胞后荧光素酶强度、S蛋白切割效率及ROS水平均降低(均P＜0.05),线粒体长度更长(P＜0.0001),MMP水平升高(P＜0.001).结论 Omicron变异株XBB.1.16-S对非小细胞肺癌细胞H1299-ACE2的感染能力和线粒体损伤能力较亲代病毒有一定程度减弱,这为肺癌合并新型冠状病毒肺炎的细胞学变化及治疗靶点的研究奠定了重要基础.
AbstractList R734.2; 目的 探讨新型冠状病毒Omicron变异株XBB.1.16刺突蛋白(XBB.1.16-S)感染人非小细胞肺癌细胞H1299-ACE2的病毒学特征及其对H1299-ACE2细胞线粒体受损的影响.方法 以亲本pCMV3-SARS-CoV-2-S质粒为模板构建XBB.1.16-S基因真核表达载体(pCMV3-XBB.1.16-S质粒)后转染H1299-ACE2细胞,比较两者诱导细胞形成合胞体的能力.利用双质粒系统包装SARS-CoV-2-S和XBB.1.16-S假病毒后分别感染H1299-ACE2细胞,测定感染后宿主细胞荧光素酶强度并采用Western blot实验检测细胞中S蛋白的切割效率,采用MitoTracker Green探针观察染色细胞的线粒体形态,采用流式细胞仪检测细胞的线粒体膜电位(mitochondrial membrane potential,MMP)水平和活性氧(reactive oxygen species,ROS)水平.结果 与亲本pCMV3-SARS-CoV-2-S质粒相比,pCMV3-XBB.1.16-S质粒的T19I、V83A、G142D等关键氨基酸突变位点突变成功,且转染H1299-ACE2细胞后所形成的合胞体面积明显减小(P＜0.0001).与SARS-CoV-2-S相比,XBB.1.16-S感染H1299-ACE2细胞后荧光素酶强度、S蛋白切割效率及ROS水平均降低(均P＜0.05),线粒体长度更长(P＜0.0001),MMP水平升高(P＜0.001).结论 Omicron变异株XBB.1.16-S对非小细胞肺癌细胞H1299-ACE2的感染能力和线粒体损伤能力较亲代病毒有一定程度减弱,这为肺癌合并新型冠状病毒肺炎的细胞学变化及治疗靶点的研究奠定了重要基础.
Abstract_FL Objective To investigate the virological characteristics of human non-small cell lung cancer cell line H1299-ACE2 infected with Omicron variant XBB.1.16 spike protein(XBB.1.16-S)and its effect on mitochondrial damage in H1299-ACE2 cells.Methods The XBB.1.16-S gene eukaryotic expression vector(pCMV3-XBB.1.16-S plasmid)was constructed by using the parental pCMV3-SARS-CoV-2-S plasmid as a template and transfected into H1299-ACE2 cells,the ability of inducing syncytial formation was compared.The H1299-ACE2 cells were infected with SARS-CoV-2-S and XBB.1.16-S pseudoviruses after packaging with a double plasmid system,respectively.The luciferase intensity of the host cells after infection was determined.The cleavage efficiency of S protein in the cells was detected by Western blot.MitoTracker green probe was used to observe mitochondrial morphology of stained cells.The levels of mitochondrial membrane potential(MMP)and reactive oxygen species(ROS)of the cells were detected by flow cytometry.Results Compared with the parental pCMV3-SARS-CoV-2-S plasmid,key amino acid mutation sites such as T19I,V83A,and G142D in the pCMV3-XBB.1.16-S plasmid were successfully mutated,and the area of the syncytium formed after transfection into H1299-ACE2 cells was significantly reduced(P＜0.0001).Compared with those of SARS-CoV-2-S,the luciferase intensity,S protein cleavage efficiency and ROS level of H1299-ACE2 cells after being infected with XBB.1.16-S were decreased(all P＜0.05),while mitochondrial length was longer(P＜0.0001),the MMP level was increased(P＜0.001).Conclusions The infectivity and mitochondrial damage ability of Omicron variant XBB.1.16-S on non-small cell lung cancer H1299-ACE2 cells were weakened to some extent compared with the parental virus.This finding lays an im-portant foundation for the study of cellular changes and treatment targets in lung cancer complicated by COVID-19.
Author 徐小洁
亢小峰
孙丽颖
薛春源
潘露
陈佳欣
房廖鑫
杜祎萌
汤传昊
AuthorAffiliation 100071 北京 中国人民解放军军事科学院军事医学研究院
AuthorAffiliation_xml – name: 100071 北京 中国人民解放军军事科学院军事医学研究院
Author_FL FANG Liaoxin
XUE Chunyuan
DU Yimeng
SUN Liying
PAN Lu
KANG Xiaofeng
XU Xiaojie
CHEN Jiaxin
TANG Chuanhao
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  fullname: TANG Chuanhao
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  fullname: SUN Liying
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DocumentTitle_FL Effects of Omicron variant XBB.1.16 spike protein on biological function of human non-small cell lung cancer cells
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Keywords Mitochondrion
H1299-ACE2细胞
Non-small cell lung cancer
XBB.1.16
Omicron
线粒体
非小细胞肺癌
H1299-ACE2 cells
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PublicationTitle 中国癌症防治杂志
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Snippet R734.2; 目的 探讨新型冠状病毒Omicron变异株XBB.1.16刺突蛋白(XBB.1.16-S)感染人非小细胞肺癌细胞H1299-ACE2的病毒学特征及其对H1299-ACE2细胞线粒体受损的影响.方法...
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StartPage 271
Title Omicron变异株XBB.1.16刺突蛋白对人非小细胞肺癌细胞的生物学功能的影响
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