基于缺氧相关基因的头颈部鳞状细胞癌分子分类及STC2的临床意义  

Molecular classification of head and neck squamous cell carcinoma based on hypoxia-related genes and clinical significance of STC2

作　　者：朱嘉宁 王天天 张瑞[2] 宋红权[3] ZHU Jianing;WANG Tiantian;ZHANG Rui;SONG Hongquan(School of Stomatology,Harbin Medical University,Harbin 150000,China;Department of Stomatology,Nangang Branch of Heilongjiang Province Hospital,Harbin 150000,China;Department of Oral Maxillofacial,The First Affiliated Hospital of Harbin Medical University,Harbin 150000,China)

机构地区：[1]哈尔滨医科大学口腔医学院,黑龙江哈尔滨150000 [2]黑龙江省医院南岗院区口腔科,黑龙江哈尔滨150000 [3]哈尔滨医科大学附属第一医院口腔颌面外科,黑龙江哈尔滨150000

出　　处：《口腔疾病防治》2025年第5期345-358,共14页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：黑龙江省自然科学基金项目(LH2022H0569);黑龙江省博士后科研启动基金(LBH-Q21143)。

摘　　要：目的基于缺氧相关基因(hypoxia-related genes,HRGs)表达谱构建头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)分类系统,并探讨缺氧基因斯钙素2(stanniocalcin 2,STC2)的临床病理特征及生物学功能。方法从癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中获取546例HNSCC样本的转录组数据及临床信息,基于200个HRGs的表达谱,采用非负矩阵分解(non-negative matrix factoriza-tion,NMF)识别HNSCC亚类,通过比较亚类间的肿瘤突变负荷、功能富集分析、药物敏感性及临床特征,评估各亚类的分子特征及预后差异。采用LASSO-Cox回归筛选预后相关基因并构建预后模型。利用TCGA数据库中口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)相关数据,分析STC2在OSCC与对照样本中的表达差异,利用qRT-PCR和免疫组化检测OSCC样本中STC2的mRNA和蛋白表达;在CAL-27细胞中敲低STC2,通过qRT-PCR和Western blot验证敲低效率,并通过CCK-8实验和细胞划痕试验评估STC2对细胞增殖和迁移能力的影响。结果基于HRGs表达谱,HNSCC被分为了3个亚类(C1、C2、C3)。C1亚类缺氧活性中等,预后良好;C2亚类缺氧活性最高,预后差,且对CTLA-4抑制剂敏感(P<0.05);C3亚类缺氧活性最低,预后中等,STC2属于C3亚类。HNSCC中细胞周期依赖性激酶抑制剂2A(cyclin-dependent kinase inhibitor 2A,CDKN2A)、磷脂酰肌醇-4,5-二磷酸3-激酶催化亚基α(phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha,PIK3CA)和肿瘤蛋白p53(tumor protein p53,TP53)突变频率较高,C1亚类的基因组增益和缺失负担显著高于C3亚类(P<0.05),而C2亚类的基因组增益显著高于C3亚类(P<0.05)。C2亚类显著富集于甘氨酸代谢、碱基切除修复等缺氧相关通路(P<0.05),提示其独特的分子特征。C1、C2和C3亚类在性别(男性)(Cramer's V=0.15)、辐射暴露(Cramer's V=0.12)、药物治疗(Cramer's V=0.18)及病理分级(G1/G2)(Cramer's V=0.25)方面均呈显著正�Objective To construct a molecular classification system for head and neck squamous cell carcinoma(HNSCC)utilizing hypoxia-related gene(HAG)expression profiles,and to comprehensively examine the clinicopathological significance and biological functions of the hypoxia gene stanniocalcin 2(STC2)in HNSCC.Methods Transcriptomic data and clinical information of 546 HNSCC samples were obtained from The Cancer Genome Atlas(TCGA)database,and based on the expression profiles of 200 HRGs,HNSCC was classified subclasses using non-negative matrix factorization(NMF).HNSCC was classified into three subclasses(C1,C2,and C3),and the molecular characteristics and prognostic differences of the subclasses were assessed by comparing the tumor mutation load,functional enrichment analysis,drug sensitivity,and clinical features among the subclasses.LASSO-Cox regression was used to screen prognosis-related genes and construct prognostic models.Using oral squamous cell carcinoma(OSCC)-related data in the TCGA database,we analyzed the expression differences of STC2 in OSCC and control samples,and detected the mRNA and protein expression of STC2 in oral squamous carcinoma samples using qRT-PCR and immunohistochemistry.We knocked down STC2 in CAL-27 cells and verified the knockdown efficiency by qRT-PCR and Western blot.CCK-8 assay and cell scratch assay were used to assess the effect of STC2 on cell proliferation and migration ability.Results Based on HRGs expression profiles,HNSCC was categorized into three subclasses(C1,C2,and C3).Subclass C1 had moderate hypoxic activity and good prognosis;subclass C2 had the highest hypoxic activity,poor prognosis,and poor sensitivity to CTLA-4 inhibitors(P<0.05);subclass C3 had the lowest hypoxic activity and moderate prognosis,and STC2 belonged to subclass C3.The frequency of cyclin-dependent kinase inhibitor 2A(CDKN2A),phosphatidylinositol4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA),and tumor protein p53(TP 53)mutations was higher in HNSCC.C1 genomic gain and deletion burden were signif

关 键 词：头颈部鳞状细胞癌 口腔鳞状细胞癌 缺氧相关基因 癌症基因组图谱 生物信息学分析 斯钙素2 预后标记 预后模型 肿瘤微环境 免疫治疗 

分 类 号：R78[医药卫生—口腔医学]