微RNA-133b通过靶向转胶蛋白-2调控非小细胞肺癌增殖及侵袭转移能力的机制研究  

Study on the mechanism of miR-133b regulating proliferation,invasion and metastasis of non-small cell lung cancer by targeting transgelin-2

作　　者：马玲[1] 李应龙 岳娜[2] 杨丽菲[1] Ma Ling;Li Yinglong;Yue Na;Yang Lifei(Department of Pulmonary Medicine(Inpatient Area 1),The Affiliated Cancer Hospital of Xinjiang Medical University,Urumqi 830011,Xinjiang,China;Department of Pathology,The Affiliated Cancer Hospital of Xinjiang Medical University,Urumqi 830011,Xinjiang,China)

机构地区：[1]新疆医科大学附属肿瘤医院肺内科(一病区),新疆乌鲁木齐830011 [2]新疆医科大学附属肿瘤医院病理科,新疆乌鲁木齐830011

出　　处：《肿瘤综合治疗电子杂志》2025年第1期114-123,共10页Journal of Multidisciplinary Cancer Management(Electronic Version)

基　　金：新疆维吾尔自治区自然科学基金面上项目(2022D01C295)。

摘　　要：目的探究微RNA-133b(microRNA-133b,miR-133b)通过靶向转胶蛋白-2(transgelin-2,TAGLN2)调控非小细胞肺癌(non-small cell lung cancer,NSCLC)增殖及侵袭转移能力的机制。方法收集2021年1月至2022年12月于新疆医科大学附属肿瘤医院经手术切除的86例NSCLC患者的肿瘤样本与对应癌旁组织。对筛选的A549和NCI-H460细胞进行多组慢病毒转染实验,以研究不同基因表达[阴性对照组(转染miRNA阴性对照模拟物)、miR-133b模拟物组(转染miR-133b模拟物)、Scrambled组(转染miRNA阴性对照模拟物+pcDNA3.1空质粒)、沉默TAGLN2组(转染TAGLN2短发夹RNA)、TAGLN2组(转染TAGLN2)、miR-133b模拟物+TAGLN2组(转染miR-133b模拟物+TAGLN2)]对细胞的影响。采用实时定量反转录聚合酶链反应检测组织、细胞系miR-133b、TAGLN2表达水平,采用免疫组织化学检测组织TAGLN2表达水平,分析miR-133b表达水平与临床病理特征的关系。利用Starbase数据库进行预测,分析miR-133b与TAGLN2之间潜在的结合位点,验证miR-133b与TAGLN2的靶向作用关系。通过细胞计数(cell counting kit-8,CCK-8)试剂盒、克隆形成实验、划痕实验、Transwell实验观察miR-133b、TAGLN2对A549、NCI-H460细胞增殖、迁移、侵袭的影响。蛋白质印记法检测TAGLN2和上皮-间充质转化(epithelial-mesenchymal transformation,EMT)相关信号通路蛋白表达。结果miR-133b在NSCLC中呈低表达(P<0.05),TAGLN2呈高表达(P<0.05)。miR-133b表达与TAGLN2水平负相关(r=-0.2195,P=0.0423),靶向调控TAGLN2表达。与阴性对照组相比,miR-133b模拟物组A549和NCI-H460细胞增殖、克隆形成、迁移、侵袭能力均显著降低(均P<0.05)。与Scramble组相比,沉默TAGLN2组A549和NCI-H460细胞增殖、克隆形成、迁移、侵袭能力、TAGLN2、磷酸化胰岛素样生长因子1受体(phospho-insulin-like growth factor 1 receptor,p-IGF1R)、磷酸化磷脂酰肌醇3激酶(phospho-phosphoinositide 3-kinase,p-PI3K)、磷酸化蛋白激酶B(phospho-proteObjective To investigate the mechanism by which microRNA-133b(miR-133b)regulates the proliferation,invasion and metastasis of non-small cell lung cancer(NSCLC)by targeting transgelin 2(TAGLN2).Method Tumor samples and corresponding adjacent tissues were collected from 86 cases of NSCLC patients who underwent surgical resection in The Affiliated Cancer Hospital of Xinjiang Medical University from January 2021 to December 2022.Multiple lentiviral transfection experiments were conducted on the screened A549 and NCI-H460 cells to investigate the effects of different gene expressions[negative control group(transfected with miRNA negative control mimic),miR-133b mimic group(transfected with miR-133b mimic),Scrambled group(transfected with miRNA negative control mimic+pcDNA3.1 empty plasmid),TAGLN2 silencing group(transfected with TAGLN2 short hairpin RNA),TAGLN2 group(transfected with TAGLN2),miR-133b mimic+TAGLN2 group(transfected with miR-133b mimic+TAGLN2)]on the cells.Real-time quantitative reverse transcription polymerase chain reaction was used to detect the expression levels of miR-133b and TAGLN2 in tissues and cell lines.Immunohistochemical staining was performed to detect the expression level of TAGLN2 in tissues.The relationship between miR-133b expression levels and clinicopathological characteristics was analyzed.Using the Starbase database,potential binding sites between miR-133b and TAGLN2 were predicted and analyzed,and the targeting relationship between miR-133b and TAGLN2 was verified.The effects of miR-133b and TAGLN2 on the proliferation,migration,and invasion of A549 and NCI-H460 cells were observed through cell counting kit-8(CCK-8)assays,clone formation experiments,wound healing assays,and Transwell experiments.Western blot analysis was conducted to detect the expression of TAGLN2 and proteins related to the epithelial-mesenchymal transformation(EMT)signaling pathway.Result miR-133b was lowly expressed,while TAGLN2 was highly expressed in NSCLC(P<0.05).The expression of miR-133b was negatively co

关 键 词：非小细胞肺癌 微RNA-133b 转胶蛋白-2 增殖 侵袭 

分 类 号：R73[医药卫生—肿瘤]