去泛素化酶PSMD7对喉鳞状细胞癌恶性进展调控的作用研究

Effect of deubiquitinating enzyme PSMD7 on regulation of laryngeal cancer malignant progression

  • 摘要:
    背景 PSMD7作为去泛素化酶家族中的重要一员,在癌症的发生、发展中起着至关重要的作用,但其在喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)中的作用研究较少。目的 探讨去泛素化酶PSMD7在LSCC中的临床意义,明晰其与患者预后的关联,剖析其免疫浸润特性及药物敏感性,同借助体外细胞实验,对PSMD7影响LSCC细胞生物学功能的作用进行验证。方法 运用TCGA数据库结合R语言工具进行生物信息学分析,包括表达差异分析、生存分析、富集分析、免疫浸润分析以及药物敏感性分析。采用喉鳞癌细胞系开展体外实验,通过质粒转染和siRNA技术构建PSMD7过表达组(OE-PSMD7)、敲减组(si-PSMD7)及对照组(Control/si-NC)。利用实时荧光定量 PCR(quantitative real-time PCR,qPCR)及 Western blot 实验检测 PSMD7 在喉癌细胞中的表达水平;通过 CCK-8 实验及克隆形成实验评估细胞增殖,Transwell、划痕实验检测细胞迁移与侵袭;Western blot分析细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)。结果 差异分析发现PSMD7在喉癌组织中表达上调(P<0.001)。预后分析显示,高表达组喉癌患者总生存期低于低表达组(P<0.001),且 PSMD7 表达与 LSCC 患者的临床分期及 T 分期相关。富集分析表明,PSMD7 通过 ATM 等信号通路影响LSCC恶性进展。免疫浸润分析发现PSMD7Th2细胞等存在正相关关系,而与B细胞等存在负相关关系。PSMD7与达拉非尼、雷帕霉素等药物的敏感性相关。PSMD7能够促进LSCC细胞增殖(P<0.05)、迁移(P<0.05)与侵袭(P<0.05),且PSMD7能够调控EMT相关蛋白促进喉鳞状细胞癌细胞发生EMT。结论 去泛素化酶PSMD7表达水平与患者的预后状况、肿瘤免疫浸润特征、化疗药物的选择及疗效紧密相关,可对喉鳞状细胞癌的增殖、迁移侵袭及EMT作用产生影响。

     

    Abstract:
    Background PSMD7, a key member of the deubiquitinating enzyme family, plays a critical role in cancer development and progression. However, its role in laryngeal squamous cell carcinoma (LSCC) remains poorly understood. Objective To explore the clinical significance of deubiquitinating enzyme PSMD7 in LSCC, clarify its association with patient prognosis, analyze its immune infiltration characteristics and drug sensitivity, and verify the effect of PSMD7 on the biological functions of LSCC cells through in vitro cell experiments.Methods Data from the TCGA database were analyzed using R language for bioinformatics analyses, including differential expression analysis, survival analysis, enrichment analysis, immune infiltration analysis, and drug sensitivity analysis, to explore the impacts of PSMD7 on clinical prognosis, immune infiltration, and drug sensitivity in laryngeal cancer patients. In vitro experiments were performed using LSCC cell lines (AMC-HN-8 and FD-LSC-1). PSMD7 overexpression (OE-PSMD7), knockdown (si-PSMD7), and control (Control/si-NC) groups were established via plasmid transfection and siRNA technology. Quantitative real-time PCR (qPCR) and Western blot were used to detect PSMD7 expression in LSCC cells. CCK-8 and colony formation assays were used to evaluate cell proliferation; Cell migration and invasion was assessed by Transwell and wound-healing assays; Western blot was applied to analyze the expression of epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin, and vimentin).Results Differential expression analysis revealed that PSMD7 was significantly upregulated in laryngeal cancer tissues (P < 0.001). Survival analysis showed that patients with high PSMD7 expression had a significantly shorter overall survival (OS) than those with low expression (P < 0.001), and PSMD7 expression was correlated with clinical stage and T stage in LSCC patients. Enrichment analysis indicated that PSMD7 influenced the malignant progression of LSCC through signaling pathways including ATM. Immune infiltration analysis demonstrated positive correlations between PSMD7 expression and Th2 cells, eosinophils, and macrophages, while negative correlations were observed with B cells and CD8+ T cells. Drug sensitivity analysis further showed associations between PSMD7 expression and sensitivity to chemotherapeutic agents such as dabrafenib and rapamycin.In vitro functional experiments revealed that PSMD7 promoted LSCC cell proliferation (P < 0.05), migration (P < 0.05), and invasion (P < 0.05). Western blot analysis showed that PSMD7 regulated the expression of epithelial-mesenchymal transition (EMT) -related proteins (E-cadherin, N-cadherin, vimentin), thereby facilitating EMT in laryngeal squamous cell carcinoma cells. Conclusion The expression level of the deubiquitinating enzyme PSMD7 is closely related to the prognosis of patients, the characteristics of tumor immune infiltration, the selection of chemotherapeutic drugs, and their efficacy. It can affect the proliferation, migration, invasion, and EMT of laryngeal squamous cell carcinoma.

     

/

返回文章
返回