基于Fibulin-5表达探讨甲状腺癌血管新生调节的分子机制  

Molecular mechanism of fibulin-5 expression in the regulation of angiogenesis in thyroid cancer

作　　者：徐志诚[1] 李春海[1] 吕宗峻 王晓东[1] XU Zhi-cheng;LI Chun-hai;LYU Zong-jun;WANG Xiao-dong(Department of General Surgery,Jiamusi Central Hospital,Jiamusi 154002,Heilongjiang,China)

机构地区：[1]佳木斯市中心医院普外二科,黑龙江佳木斯154002

出　　处：《广东医学》2025年第1期41-47,共7页Guangdong Medical Journal

基　　金：黑龙江省卫生健康科研基金项目(2022H03211)。

摘　　要：目的探讨甲状腺癌组织微环境中Fibulin-5(FBLN5)在血管新生和血管透性调节中的分子机制。方法分析2022年6至12月在佳木斯市中心医院诊断为甲状腺乳头状癌(PTC)的10个匹配的PTC组织和邻近的正常甲状腺组织样本。使用蛋白质印迹法、RT-qPCR分析样本内皮细胞(ECs)中FBLN5表达。将人真皮微血管ECs(HDMECs)和甲状腺癌细胞系KTC-1进行了接触共培养实验,以考察HDMECs中FBLN5的表达变化。为了研究FBLN5在调节ECs血管生成中的功能,将HDMECs分为载体(Vector)组、FBLN5过表达质粒(FBLN5)组、随机序列(sh-NC)组、FBLN5敲低慢病毒(sh-FBLN5)组。分别用FBLN5过表达质粒或shRNA慢病毒上调或敲低HDMECs细胞中FBLN5,通过WST-1分析、管形成试验、Transwell试验评估FBLN5过表达或敲低对细胞的增殖能力、管形成、迁移影响。通过向NOD-SCID小鼠的侧翼注射与Vector或FBLN5转染的HDMECs混合的PTC细胞来进行侧翼肿瘤模型,通过免疫组化检测肿瘤组织中微血管密度。结果与正常ECs相比,PTC患者的肿瘤相关ECs中FBLN5 mRNA和蛋白表达均显著下调(P<0.05)。FBLN5在与和不与KTC-1细胞共培养的HDMECs中表达相当。与Vector组相比,FBLN5转染的HDMECs在孵育48 h、72 h时的细胞活力显著降低(P<0.05)。与sh-NC组相比,sh-FBLN5转染的HDMECs在孵育48、72 h时的细胞活力显著增加(P<0.05)。与Vector组相比,FBLN5转染的HDMECs的细胞迁移数(P<0.001)和成管数(P<0.001)显著降低;与sh-NC组相比,sh-FBLN5转染的HDMECs的细胞迁移数(P=0.005)和成管数(P<0.001)显著增加。FBLN5转染HDMECs在第3、6、10和14天抑制了PTC肿瘤的生长。FBLN5组肿瘤组织中CD31标记的微血管密度显著低于Vector组(P=0.005),和FBLN5蛋白表达显著高于Vector组(P<0.001)。结论缺氧诱导的内皮细胞中HIF1α上调通过抑制FBLN5表达来促进PTC血管生成。Objective To investigate the molecular mechanism of Fibulin-5(FBLN5)in the regulation of angiogenesis and vascular permeability in the tumor microenvironment of thyroid cancer.Methods Ten matched samples of papillary thyroid carcinoma(PTC)tissues and adjacent normal thyroid tissues diagnosed from June 2022 to December 2022 at Jiamusi Central Hospital were analyzed.Protein immunoblotting and RT-qPCR were used to assess FBLN5 expression in endothelial cells(ECs).Co-culture experiments with human dermal microvascular ECs(HDMECs)and thyroid cancer cell line KTC-1 were performed to investigate changes in FBLN5 expression in HDMECs.In order to study the function of FBLN5 in regulating the angiogenesis of ECs,HDMECs were divided into Vector group,FBLN5 overexpression plasmid group,random sequence(sh-NC)group and FBLN5 knock-down lentivirus(sh-FBLN5)group.Overexpression plasmids or shRNA lentivirus targeting FBLN5 were used to upregulate or downregulate FBLN5 in HDMECs.Cell proliferation,tube formation,and migration were assessed using WST-1,tube formation,and Transwell assays,respectively.A xenograft tumor model was established in NOD-SCID mice by injecting PTC cells mixed with HDMECs transfected with Vector or FBLN5.Immunohistochemistry was performed to assess microvessel density in tumor tissues.Results Compared to normal ECs,FBLN5 mRNA and protein expression in tumor-associated ECs of PTC patients were significantly downregulated(P<0.05).FBLN5 expression in HDMECs co-cultured with and without KTC-1 cells was comparable.Compared to the Vector group,the cell viability of FBLN5-transfected HDMECs was significantly reduced at 48 h and 72 h(P<0.05).Compared to the sh-NC group,the cell viability of sh-FBLN5-transfected HDMECs was significantly increased at 48 h and 72 h(P<0.05).The cell migration(P<0.001)and tube formation(P<0.001)of FBLN5-transfected HDMECs were significantly decreased compared to the Vector group;compared to the sh-NC group,the cell migration(P=0.005)and tube formation(P<0.001)of sh-FBLN5-transfected HDM

关 键 词：FIBULIN-5 甲状腺癌 血管新生 缺氧 

分 类 号：R581[医药卫生—内分泌] R736[医药卫生—内科学]