鞘氨醇激酶1通过TGF-β依赖性EMT参与口腔黏膜下纤维化恶性转化  

Sphingosine kinase 1 is involved in the malignant transformation of oral submucosal fibrosis through TGF-β-dependent EMT

作　　者：尤轶阳 王宗康 谭劲[3] 肖艳波[3] YOU Yiyang;WANG Zongkang;TAN Jin;XIAO Yanbo(School of Graduate,Hunan University of Traditional Chinese Medicine,Changsha Hunan 410208;Department of Stomatology,The People′s Hospital of Liuyang,Liuyang Hunan 410300;Department of Stomatology,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410000,China)

机构地区：[1]湖南中医药大学研究生院,湖南长沙410208 [2]湖南省浏阳市人民医院口腔科,410300 [3]湖南中医药大学第一附属医院口腔科,湖南长沙410000

出　　处：《蚌埠医科大学学报》2025年第3期287-292,共6页Journal of Bengbu Medical University

基　　金：国家自然科学基金项目(81874496)。

摘　　要：目的:探讨鞘氨醇激酶1(SphK1)在槟榔碱相关口腔黏膜下纤维化发病机制中的功能作用及其相关机制。方法:通过槟榔碱刺激人口腔黏膜成纤维细胞(BMF)建立体外模型,并使用SphK1特异性抑制剂PF-543和转染SphK1过表达质粒(SphK1)、Smad2/3敲低(shSmad2/3)进行处理。通过蛋白质印迹、免疫荧光染色分析细胞上皮-间质转化(EMT)标志物表达,通过伤口愈合试验、Transwell侵袭试验检测细胞的迁移和侵袭能力。采用基因集富集分析法分析BMF中SphK1调节的潜在途径。结果:槟榔碱以时间和浓度依赖性方式增加BMF中SphK1的表达(P<0.05)。与对照组相比,槟榔碱组迁移和侵袭的细胞数量增加(P<0.05),并且BMF上皮标志物E-cadherin表达下调而间充质标志物Vimentin表达上调(P<0.05);PF-543组细胞迁移和侵袭的数量较槟榔碱组降低(P<0.05),并增加了BMF中的上皮标志物E-cadherin表达而降低了间充质标志物Vimentin表达(P<0.05)。基因集富集分析结果显示,转化生长因子-β(TGF-β)信号传导是SphK1过表达时最显著激活的途径。Smad2/3敲低有效地阻断了SphK1诱导的细胞迁移和侵袭(P<0.05),并逆转SphK1过表达细胞中下游EMT标志物的表达(P<0.05)。结论:SphK1通过TGF-β依赖性EMT参与口腔黏膜下纤维化恶性转化。Objective:To explore the role of sphingosine kinase 1(SphK1)in the pathogenesis of arecoline-related oral submucosal fibrosis and its related mechanisms.Methods:An in vitro model of human oral mucosal fibroblasts(BMF)stimulated by arecoline was established,and treated with SphK1 specific inhibitor PF-543,SphK1 overexpression plasmid(SphK1)and Smad2/3 knockdown(shSmad2/3).The expression of epithelial-mesenchymal transition(EMT)markers in cells was detected by Western blotting and immunofluorescence staining,and the migration and invasion abilities of cells were examined by wound healing assay and Transwell invasion assay.The potential pathway of sphK1 regulation in BMF was analyzed by Gene Set Enrichment Analysis.Results:Arecoline increased the expression of SphK1 in BMF in a time-and concentration-dependent manner(P<0.05).Compared with the control group,the number of migrated and invaded cells in the arecoline group was increased(P<0.05),and the expression of epithelial marker E-cadherin was down-regulated while the expression of mesenchymal marker Vimentin was up-regulated in BMF(P<0.05).The number of migrated and invaded cells in the PF-543 group was lower than that in the arecoline group(P<0.05),and the epithelial marker E-cadherin were increased while the expression of mesenchymal marker Vimentin was decreased in BMF(P<0.05).Gene Set Enrichment Analysis showed that transforming growth factor-β(TGF-β)signaling was the most significantly activated pathway upon SphK1 overexpression.Smad2/3 knockdown effectively blocked SphK1-induced cell migration and invasion(P<0.05),and reversed the expression of downstream EMT markers in SphK1-overexpressing cells(P<0.05).Conclusions:SphK1 is involved in the malignant transformation of oral submucosal fibrosis through TGF-β-dependent EMT.

关 键 词：口腔黏膜下纤维化 鞘氨醇激酶1 上皮-间质转化 转化生长因子-Β 

分 类 号：R735.2[医药卫生—肿瘤]