LINC00973通过hsa-miR-150-5p/ABCG5轴调控非小细胞肺癌多药耐药  

LINC00973 regulates multidrug resistance of non-small-cell lung cancer through hsa-miR-150-5p/ABCG5 axis

作　　者：夏云秀 董洪亮[1,2] 刘翠兰 王飞 崔冰洁 陈微微 杜静 XIA Yunxiu;DONG Hongliang;LIU Cuilan;Wang Fei;CUI Bingjie;CHEN Weiwei;DU Jing(Department of Respiratory and Critical Care Medicine,Binzhou Medical University Hospital,Binzhou 256600,China;Medical Research Center,Binzhou Medical University Hospital,Binzhou 256600,China;Department of gynecology,Bin-zhou Medical University Hospital,Binzhou 256600,China;Department of Oncology,Binzhou Medical University Hospi-tal,Binzhou 256600,China)

机构地区：[1]滨州医学院附属医院呼吸与危重症科,山东滨州256600 [2]滨州医学院附属医院医学研究中心,山东滨州256600 [3]滨州医学院附属医院妇科,山东滨州256600 [4]滨州医学院附属医院肿瘤科,山东滨州256600

出　　处：《中国病理生理杂志》2025年第3期433-443,共11页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31900441,No.82373097);山东省自然科学基金资助项目(No.ZR2019MC026,No.22QH192);山东省中医药科技项目(No.Z20244106)。

摘　　要：目的:探讨长链非编码RNA LINC00973在非小细胞肺癌(non-small-cell lung cancer,NSCLC)多药耐药中的作用及分子机制。方法:GEPIA数据库分析LINC00973在NSCLC中的表达水平及其与患者预后的关系,RT-qPCR检测永生化支气管上皮细胞和不同NSCLC细胞系及化疗耐药NSCLC细胞中LINC00973的表达水平。构建LINC00973过表达或敲减的NSCLC细胞系,采用细胞划痕、Transwell及干性成球实验检测LINC00973对NSCLC细胞迁移侵袭能力和干性的影响;CCK-8实验检测LINC00973过表达或敲减后,化疗药和靶向药对NSCLC细胞半数抑制浓度的变化。RNA-seq分析LINC00973过表达后靶基因簇改变;LncBase Predicted v.2和TargetScan数据库分析LINC00973与靶基因共同结合的微小RNA(microRNA,miRNA);合成并转染目标miRNA-mimic/inhibitor至LINC00973过表达或敲减的NSCLC细胞中,RT-qPCR和Western blot实验检测LINC00973表达水平及其靶基因的mRNA和蛋白表达水平的变化。结果:GEPIA数据库分析结果显示,LINC00973在肺腺癌和肺鳞癌中表达上调,且与NSCLC患者不良预后相关;不同NSCLC细胞系及NSCLC化疗耐药细胞(A549/DDP和A549/5-FU细胞)中LINC00973表达上调。过表达LINC00973后A549和H520细胞的迁移侵袭能力和干性明显增强,对化疗药和靶向药的敏感性明显下降;敲减LINC00973后A549和H520细胞的迁移侵袭能力和干性明显减弱,对化疗药和靶向药的敏感性明显增强。RNA-seq结合RT-qPCR实验结果显示,过表达LINC00973的A549和H520细胞中,ATP结合盒转运蛋白G5(ATP-binding cassette transporter G5,ABCG5)表达上调,ABC转运体通道被激活。数据库分析结合细胞实验证实LINC00973通过竞争性结合hsa-miR-150-5p调控ABCG5的表达。结论:LINC00973通过与ABCG5竞争性结合hsa-miR-150-5p的方式,上调ABCG5的表达,进而激活ABC转运体通道并促进抗肿瘤药物外排,导致NSCLC产生多药耐药。AIM:This study aims to investigate the mechanism by which LINC00973 regulates multidrug resistance of non-small-cell lung cancer(NSCLC).METHODS:The GEPIA database was employed to analyze the expression levels of LINC00973 in NSCLC and its correlation with patient prognosis in clinical settings.RT-qPCR was utilized to assess LINC00973 expression in various NSCLC cell lines and chemoresistant cells.The migration,invasion,stemness capacity,and drug sensitivity of NSCLC cells with either overexpression or knockdown of LINC00973 were evaluated using wound healing assay,Transwell assay,sphere formation assay,and CCK-8 assay.RNA sequencing was conducted on LINC00973-overexpressing and parental NSCLC cells to identify dysregulated pathways and targets.The LncBase Predicted v.2 and TargetScan databases were used to predict the microRNAs(miRNAs)co-bound by LINC00973 and their target genes.Mimics and inhibitors of candidate miRNAs were synthesized and transfected into NSCLC cells subjected to LINC00973 overexpression or knockdown.Changes in the expression level of LINC00973,and the mRNA and protein expression levels of its target genes were assessed using RT-qPCR and Western blot.RESULTS:Analysis of the GEPIA database revealed that LINC00973 was significantly up-regulated in lung adenocarcinoma and lung squamous cell carcinoma,correlating with poor prognosis of NSCLC patients.The LINC00973 expression was elevated in various NSCLC and chemoresistant cell lines(A549/DDP and A549/5-FU).Overexpression of LINC00973 in A549 and H520 cells markedly enhanced their migration,invasion,and stemness capabilities,while concurrently reducing sensitivity to chemotherapy and targeted therapies.RNA sequencing,along with RT-qPCR results,indicated that ATP-binding cassette transporter G5(ABCG5)was activated in LINC00973-overexpressing A549 and H520 cells.Further database analyses and dual transfection experiments confirmed that LINC00973 regulated ABCG5 expression through competitive binding with hsa-miR-150-5p.CONCLUSION:LINC00973,which is a

关 键 词：非小细胞肺癌 多药耐药 LINC00973 微小RNA-150-5p ATP结合盒转运蛋白G5 

分 类 号：R734.2[医药卫生—肿瘤] R318.0[医药卫生—临床医学] Q756[生物学—分子生物学]