摘要
背景与目的靶向治疗在肺鳞癌中的效果不理想,而免疫治疗较低的应答率阻碍了其在肺鳞癌中的应用,因此,探索肺鳞癌治疗的新策略十分迫切。铁死亡在抑制肿瘤方面发挥了重要作用,本研究旨在探究靶向3-羟基-3-甲基戊二酰辅酶A合酶1(3-hydroxy-3-methylglutaryl-Co A synthase 1,HMGCS1)对肺鳞癌细胞铁死亡的调控和作用机制,为肺鳞癌治疗提供新的研究思路和方向。方法通过肿瘤基因组图谱(e Cancer Genome Atlas,TCGA)和临床蛋白质组肿瘤分析联盟(Clinical Proteomic Tumor Analysis Consortium,CPTAC)在线蛋白数据库分析HMGCS1在肺鳞癌中的表达情况;通过Kaplan-Meier Plotter在线生存数据库分析HMGCS1与肺癌生存时间的关系;通过免疫组化验证HMGCS1在肺鳞癌组织的表达水平;在肺鳞癌细胞系SKMES细胞通过小干扰RNA(small interfering RNA,si RNA)干扰HMGCS1表达后,通过CCK8和Transwell检测细胞活性和细胞迁移能力;通过流式细胞术检测干扰HMGCS1后或用HMGCS1抑制剂hymeglusin处理后细胞凋亡水平;通过流式和高内涵共聚焦荧光成像系统分别检测抑制HMGCS1后SKMES细胞中Fe~(2+)、活性氧(reactive oxygen species,ROS)和脂质过氧化水平;通过Western blot检测抑制HMGCS1后铁死亡通路标志蛋白ACSL4、GPX4和SLC7A11表达。结果HMGCS1 m RNA和蛋白水平在肺鳞癌均显著高表达;si RNA干扰HMGCS1表达抑制了肺鳞癌细胞增殖活性和迁移能力,但对细胞凋亡没有显著影响。干扰HMGCS1后或用HMGCS1抑制剂hymeglusin处理后显著促进了SKMES细胞内Fe~(2+)、ROS和脂质过氧化水平,促进肺鳞癌细胞铁死亡;Western blot检测显示抑制HMGCS1显著促进了ACSL4的表达。结论抑制肺鳞癌靶点HMGCS1可促进肺癌细胞铁死亡,为肺鳞癌筛选新的治疗靶点提供了研究基础。
Background and objective Targeted therapies are ineffective in lung squamous cancer(LUSC),and the low response rate of immunotherapy hampers its application in LUSC,so it is urgent to explore new strategies for LUSC treatment.Ferroptosis plays an important role in tumour suppression.The aim of this study was to investigate the role and mechanism of targeting 3-hydroxy-3-methylglutaryl-CoA synthase 1(HMGCS1)in regulating ferroptosis in LUSC cells,in order to provide a new research direction for LUSC therapy.Methods The expression of HMGCS1 in LUSC was analysed by The Cancer Genome Atlas(TCGA)and Clinical Proteomic Tumor Analysis Consortium(CPTAC)online databases;the relationship between HMGCS1 and survival time of lung cancer was analysed by the Kaplan-Meier Plotter online survival database;the expression level of HMGCS1 in LUSC tissues was verified by immunohistochemistry.After interfering with HMGCS1 expression by small interfering RNA(siRNA),cell activity and cell migration ability were detected by CCK8 and Transwell assay;apoptosis was detected by flow cytometry after interfering with HMGCS1 or after treatment with the HMGCS1 inhibitor of hymeglusin;Fe2+,reactive oxygen species(ROS)and lipid peroxidation levels were detected by flow cytometry and highcontent confocal fluorescence imaging systems,respectively in SKMES cells after inhibition of HMGCS1;and Western blot was performed to detect the expression of ACSL4,GPX4 and SLC7A11,which are markers of the ferroptosis pathway after inhibition of HMGCS1.Results HMGCS1 mRNA and protein levels were significantly high in LUSC;siRNA interference with HMGCS1 expression inhibited the proliferative activity and migration ability of LUSC cells,but had no significant effect on apoptosis.Interference with HMGCS1 or treatment with the HMGCS1 inhibitor of hymeglusin significantly promoted intracellular Fe2+,ROS and lipid peroxidation levels in SKMES cells,and induced ferroptosis in LUSC cells;Western blot assay showed that inhibition of HMGCS1 significantly promoted the expr
作者
倪银芸
杨瑛
张立
Yinyun NI;Ying YANG;Li ZHANG(Institute of Respiratory Health,Frontiers Science Center for Disease-related Molecular Network,West China Hospital,Sichuan University,Chengdu 610000,China)
出处
《中国肺癌杂志》
CAS
CSCD
北大核心
2024年第5期330-336,共7页
Chinese Journal of Lung Cancer
基金
国家自然科学基金项目(No.82173251)资助。
