摘要
研究微小RNA-21(microRNA-21,miR-21)通过核因子κB(nuclear factor-κB,NF-κB)信号通路对甲状腺乳头状癌(papillary thyroid carcinoma,PTC)细胞侵袭转移的调控作用及分子机制。收集PTC患者癌组织及癌旁组织,通过实时荧光定量PCR(qRT-PCR)检测miR-21表达水平。构建miR-21过表达PTC细胞株(TPC-1和BCPAP),利用Transwell实验分析细胞迁移及侵袭能力变化,Western blot检测NF-κB通路关键蛋白(p65、IκBα)及上皮-间质转化(epithelial-mesenchymal transition,EMT)标志物、基质金属蛋白酶-2(matrix metalloproteinase 2,MMP-2)表达。与癌旁正常组织相比,miR-21在PTC组织中的表达显著低于正常组织(P<0.01)。进一步分析表明,该microRNA的低表达状态与患者淋巴结转移发生率呈统计学显著的负向关联。过表达miR-21可显著抑制PTC细胞迁移及侵袭能力,同时减弱NF-κB信号通路活性,表现为细胞质IκBα水平升高、核内p65蛋白积累减少及下游MMP-2表达下调。此外,miR-21通过促进IκBα蛋白稳定性抑制p65核转位,进而阻断EMT进程。miR-21通过抑制NF-κB信号通路削弱PTC细胞侵袭转移能力,其作用与调控IκBα/p65/MMP-2轴密切相关,提示miR-21作为抑癌因子可能成为PTC靶向治疗的潜在分子标志物。
The primary objective of this study was to investigate the regulatory role and underlying molecular mechanisms of microRNA21(miR-21)in modulating the invasion and metastasis of papillary thyroid carcinoma(PTC)cells through the nuclear factor-κB(NF-κB)signaling pathway.To achieve this,the authors collected paired clinical tissue samples,including 33 cases of PTC tumors and adjacent normal tissues from patients diagnosed between February 2024 and September 2024,and analyzed miR-21 expression levels using quantitative real-time PCR(qRT-PCR).Clinical pathological characteristics,such as age,gender,tumor size,lymph node metastasis status,and TNM staging,were correlated with miR-21 expression.In vitro,two human PTC cell lines,TPC-1 and BCPAP,were transfected with miR-21 mimics using Lipofectamine 3000 to establish stable overexpression models,with transfection efficiency validated by qRT-PCR.Functional assays,including transwell migration and invasion experiments,were conducted to assess cellular behaviors,where cells were seeded in Matrigel-coated or uncoated chambers and incubated for 18 hours,followed by microscopic quantification.Western blot analysis was employed to evaluate the expression of key NF-κB pathway components(such as cytoplasmic IκBαand nuclear p65)and epithelial-mesenchymal transition(EMT)-related markers like matrix metalloproteinase-2(MMP-2),with tubulin and histone H3 serving as internal controls for total and nuclear proteins,respectively.Statistical analyses were performed using SPSS software,with data presented as x±s and significance determined by Student's t-test or ANOVA.Results demonstrated that miR-21 expression was significantly downregulated in PTC tissues compared to normal adjacent tissues(P<0.01),and this low expression was negatively correlated with lymph node metastasis incidence(P=0.033).Overexpression of miR21 markedly suppressed the migration and invasion capabilities of PTC cells;for instance,in BCPAP cells,migration and invasion cell counts decreased to 109.8±4.8 and 63.2±5.0,respectively,compared to controls(262.6±8.1 and 119.4±7.0,P<0.01).Mechanistically,miR-21 overexpression led to increased cytoplasmic IkBαprotein stability,reduced nuclear accumulation of p65,and downregulated MMP-2 expression,indicating inhibition of NF-κB pathway activation and subsequent suppression of EMT progression.These findings conclude that miR-21 acts as a tumor suppressor in PTC by attenuating invasion and metastasis through repression of the NF-κB signaling axis,specifically via the IκBα/p65/MMP-2 cascade,highlighting its potential as a therapeutic biomarker for targeted interventions in PTC management.
作者
游伟
邓修健
何英强
YOU Wei;DENG Xiujian;HE Yingqiang(The Third People's Hospital of Shunde District,Foshan City/Beijiao Hospital of Shunde District,Foshan 528311,China)
出处
《药物生物技术》
2025年第6期760-765,共6页
Pharmaceutical Biotechnology
基金
佛山市自筹经费类科技创新项目(No.2220001004418)。
