转录-蛋白质组联合分析白藜芦醇通过EMT抑制口腔鳞状细胞癌的转移机制  

Transcription-proteome analysis of the mechanism of resveratrol inhibiting the metastasis of oral squamous cell carcinoma

作　　者：杨清 王治林[1,3] 钟越 钟晓武 邱亚[5] 李丽华[1,2] YANG Qing;WANG Zhi-lin;ZHONG Yue;ZHONG Xiao-wu;QIU Ya;LI Li-hua(Department of Stomatology,Affiliated Hospital of North Sichuan Medical College,North Sichuan Medical College,Nanchong 637000,Sichuan,China;College of Stomatology,North Sichuan Medical College,Nanchong 637000,Sichuan,China;Innovation Cooperation and Exchange Center,Tohoku National University of Dentistry,Sendai 197-0804,Miyagi,Japan;Department of Clinical Laboratory,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan,China;Science and Technology Innovation Center,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan,China)

机构地区：[1]川北医学院附属医院口腔科,四川南充637000 [2]川北医学院口腔医学院,四川南充637000 [3]日本东北大学院齿学研究科创新合作交流中心,日本宫城仙台197-0804 [4]川北医学院附属医院检验科,四川南充637000 [5]川北医学院附属医院科技创新中心,四川南充637000

出　　处：《川北医学院学报》2025年第3期283-289,共7页Journal of North Sichuan Medical College

基　　金：四川省大学生创新创业科研基金项目(S202210634077);四川省南充市应用技术研究与开发专项项目(20YFZJ0090)。

摘　　要：目的:基于转录组和蛋白质组联合探讨白藜芦醇(RSV)治疗口腔鳞状细胞癌(OSCC)的分子机制。方法:体外培养口腔鳞状细胞癌HSC-3细胞并分为对照组与RSV组,对两组进行转录组和蛋白质组联合分析,筛选干预OSCC的关键靶点和相关信号通路。将细胞分为空白对照组、RSV 50μmol/L组及RSV 100μmol/L组,通过RT-qPCR和Western blot检测各组核心差异基因/蛋白,即神经钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、上皮细胞钙黏素(E-cadherin)和基质金属蛋白酶-9(MMP9)的mRNA表达水平。结果:RSV处理HSC-3细胞后,转录组分析显示,RSV组共有422个差异表达基因,其中上调276个,下调146个,差异表达基因主要富集在上皮间充质转化(EMT)信号通路等。蛋白质组分析表明RSV组共有644个差异表达蛋白,其中上调342个,下调302个。通过转录组-蛋白质组联合对比,得到117个交集靶点,包括EMT信号通路中E-cadherin、N-cadherin、MMP9和Vimentin等。RT-qPCR结果表明,RSV上调E-cadherin mRNA的表达水平(P<0.05),下调N-cadherin、Vimentin和MMP9的mRNA表达水平(P<0.05),与转录组结果一致。Western blot结果表明,RSV处理后可增加E-cadherin的蛋白表达水平(P<0.05),降低N-cadherin、Vimentin和MMP9的蛋白表达水平(P<0.05),且与RSV浓度正相关(P<0.05),与蛋白质组结果一致。结论:RSV通过调节HSC-3细胞EMT信号通路分子E-cadherin、N-cadherin、MMP9和Vimentin的表达,抑制口腔鳞癌细胞的转移。Objective:To explore the molecular mechanism of resveratrol(RSV) in the treatment of oral squamous cell carcinoma(OSCC) based on the combination of transcriptome and proteome.Methods:Oral squamous cell carcinoma HSC-3 cells were cultured in vitro and divided into the control group and RSV group.Combined transcriptome and proteome analysis was performed in control group and RSV group to screen key targets and related signaling pathways for intervention in OSCC.The cells were divided into blank control group,RSV 50 μmol/L group,and RSV 100 μmol/L group,mRNA expression levels of core differential genes/proteins,namely neural cadherin(N-cadherin),Vimentin,epithelial cadherin(E-cadherin) and matrix metalloproteinase-9(MMP9)were detected by RT-qPCR and Western blot.Results:After RSV treatment of HSC-3 cells,transcriptome analysis showed that there were 422 differentially expressed genes in RSV group,of which 276 were up-regulated and 146 were down-regulated.Differentially expressed genes were mainly concentrated in epithelial mesenchymal transformation(EMT) signaling pathway.Proteomic analysis showed that there were 644 differentially expressed proteins in RSV group,of which 342 were up-regulated and 302 were down-regulated.By combining transcriptome-proteome comparisons,117 intersection targets were identified,including E-cadherin,N-cadherin,MMP9 and Vimentin in EMT signaling pathways.The results of RT-qPCR showed that RSV up-regulated the expression of E-cadherin mRNA and down-regulated the expression of N-cadherin,Vimentin and MMP9 mRNA,which was consistent with the transcriptomic results.Western blot experiment results showed that RSV treatment increased the protein expression level of E-cadherin and decreased the protein expression level of N-cadherin,Vimentin and MMP9,which was positively correlated with RSV concentration and consistent with proteomic results.Conclusion:RSV inhibits the metastasis of oral squamous cell by regulating the expression of EMT signaling pathway molecules E-cadherin,N-cadherin,MMP9 an

关 键 词：口腔鳞状细胞癌 白藜芦醇 转录组 蛋白质组 上皮间充质转化 

分 类 号：R78[医药卫生—口腔医学]