BACKGROUND Esophageal squamous cell carcinoma(ESCC)is one of the most common malignancies worldwide,and its development comprises a multistep process from intraepithelial neoplasia(IN)to carcinoma(CA).However,the crit...BACKGROUND Esophageal squamous cell carcinoma(ESCC)is one of the most common malignancies worldwide,and its development comprises a multistep process from intraepithelial neoplasia(IN)to carcinoma(CA).However,the critical regulators and underlying molecular mechanisms remain largely unknown.AIM To explore the genes and infiltrating immune cells in the microenvironment that are associated with the multistage progression of ESCC to facilitate diagnosis and early intervention.METHODS A mouse model mimicking the multistage development of ESCC was established by providing warter containing 4-nitroquinoline 1-oxide(4NQO)to C57BL/6 mice.Moreover,we established a control group without 4NQO treatment of mice.Then,transcriptome sequencing was performed for esophageal tissues from patients with different pathological statuses,including low-grade IN(LGIN),high-grade IN(HGIN),and CA,and controlled normal tissue(NOR)samples.Differentially expressed genes(DEGs)were identified in the LGIN,HGIN,and CA groups,and the biological functions of the DEGs were analyzed via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses.The CIBERSORT algorithm was used to detect the pattern of immune cell infilt-ration.Immunohistochemistry(IHC)was also conducted to validate our results.Finally,the Luminex multiplex cytokine analysis was utilized to measure the serum cytokine levels in the mice.RESULTS Compared with those in the NOR group,a total of 681541,and 840 DEGs were obtained in the LGIN,HGIN,and CA groups,respectively.Using the intersection of the three sets of DEGs,we identified 86 genes as key genes involved in the development of ESCC.Enrichment analysis revealed that these genes were enriched mainly in the keratinization,epidermal cell differentiation,and interleukin(IL)-17 signaling pathways.CIBERSORT analysis revealed that,compared with those in the NOR group,M0 and M1 macrophages in the 4NQO group showed stronger infiltration,which was validated by IHC.Serum cytokine analysis revealed that,compared with those in the NOR group,IL-1βand IL-6 were upregulated,while IL-10 was downregulated in the LGIN,HGIN,and CA groups.Moreover,the expression of the representative key genes,such as S100a8 and Krt6b,was verified in external human samples,and the results of immunohistochemical staining were consistent with the findings in mice.CONCLUSION We identified a set of key genes represented by S100a8 and Krt6b and investigated their potential biological functions.In addition,we found that macrophage infiltration and abnormal alterations in the levels of inflam-mation-associated cytokines,such as IL-1β,IL-6,and IL-10,in the peripheral blood may be closely associated with the development of ESCC.展开更多
目的探究食管鳞状上皮内瘤变发生风险的预测因素,构建列线图预测模型。方法回顾性收集2016年01月至2021年12月在扬州大学附属医院诊断为食管鳞状上皮内瘤变患者126例,以及同期于健康管理中心进行体检者344例,收集患者一般临床资料、血...目的探究食管鳞状上皮内瘤变发生风险的预测因素,构建列线图预测模型。方法回顾性收集2016年01月至2021年12月在扬州大学附属医院诊断为食管鳞状上皮内瘤变患者126例,以及同期于健康管理中心进行体检者344例,收集患者一般临床资料、血常规检测、肿瘤标志物、病变资料。确定食管鳞状上皮内瘤变的独立预测因子,构建列线图预测模型、绘制受试者操作特征(receiver operating characteristic,ROC)曲线,计算得出Harrell一致性指数(C-index)并采用Bootstrap自抽样法,对模型进行内部验证并绘制校准曲线及决策曲线。结果为构建食管上皮内瘤变的风险预测模型,经PSM法匹配后,本研究共纳入96例食管鳞状上皮内瘤变患者及96例健康对照。多因素Logistic回归分析显示,血红蛋白(HGB)计数、血小板(PLT)计数、血小板分布宽度(PDW)、NLR水平是食管鳞状上皮内瘤变的独立预测因子。根据以上四项指标构建的列线图模型曲线下面积(area under curve,AUC)为0.787。Bootstrap内部验证的C-index值为0.771,提示该食管鳞状上皮内瘤变风险预测模型的识别能力、一致性和临床净获益良好。结论基于HGB、PLT、PDW及NLR建立的列线图预测模型可有效鉴别有无食管鳞状上皮内瘤变病变人群,对诊断食管癌前病变有辅助价值。展开更多
基金Supported by National Natural Foundation of China,No.821742232019 Chinese and Western Medicine Clinical Collaborative Capacity Building Project for Major Difficult Diseases,No.2019-ZX-005。
文摘BACKGROUND Esophageal squamous cell carcinoma(ESCC)is one of the most common malignancies worldwide,and its development comprises a multistep process from intraepithelial neoplasia(IN)to carcinoma(CA).However,the critical regulators and underlying molecular mechanisms remain largely unknown.AIM To explore the genes and infiltrating immune cells in the microenvironment that are associated with the multistage progression of ESCC to facilitate diagnosis and early intervention.METHODS A mouse model mimicking the multistage development of ESCC was established by providing warter containing 4-nitroquinoline 1-oxide(4NQO)to C57BL/6 mice.Moreover,we established a control group without 4NQO treatment of mice.Then,transcriptome sequencing was performed for esophageal tissues from patients with different pathological statuses,including low-grade IN(LGIN),high-grade IN(HGIN),and CA,and controlled normal tissue(NOR)samples.Differentially expressed genes(DEGs)were identified in the LGIN,HGIN,and CA groups,and the biological functions of the DEGs were analyzed via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses.The CIBERSORT algorithm was used to detect the pattern of immune cell infilt-ration.Immunohistochemistry(IHC)was also conducted to validate our results.Finally,the Luminex multiplex cytokine analysis was utilized to measure the serum cytokine levels in the mice.RESULTS Compared with those in the NOR group,a total of 681541,and 840 DEGs were obtained in the LGIN,HGIN,and CA groups,respectively.Using the intersection of the three sets of DEGs,we identified 86 genes as key genes involved in the development of ESCC.Enrichment analysis revealed that these genes were enriched mainly in the keratinization,epidermal cell differentiation,and interleukin(IL)-17 signaling pathways.CIBERSORT analysis revealed that,compared with those in the NOR group,M0 and M1 macrophages in the 4NQO group showed stronger infiltration,which was validated by IHC.Serum cytokine analysis revealed that,compared with those in the NOR group,IL-1βand IL-6 were upregulated,while IL-10 was downregulated in the LGIN,HGIN,and CA groups.Moreover,the expression of the representative key genes,such as S100a8 and Krt6b,was verified in external human samples,and the results of immunohistochemical staining were consistent with the findings in mice.CONCLUSION We identified a set of key genes represented by S100a8 and Krt6b and investigated their potential biological functions.In addition,we found that macrophage infiltration and abnormal alterations in the levels of inflam-mation-associated cytokines,such as IL-1β,IL-6,and IL-10,in the peripheral blood may be closely associated with the development of ESCC.
文摘目的探究食管鳞状上皮内瘤变发生风险的预测因素,构建列线图预测模型。方法回顾性收集2016年01月至2021年12月在扬州大学附属医院诊断为食管鳞状上皮内瘤变患者126例,以及同期于健康管理中心进行体检者344例,收集患者一般临床资料、血常规检测、肿瘤标志物、病变资料。确定食管鳞状上皮内瘤变的独立预测因子,构建列线图预测模型、绘制受试者操作特征(receiver operating characteristic,ROC)曲线,计算得出Harrell一致性指数(C-index)并采用Bootstrap自抽样法,对模型进行内部验证并绘制校准曲线及决策曲线。结果为构建食管上皮内瘤变的风险预测模型,经PSM法匹配后,本研究共纳入96例食管鳞状上皮内瘤变患者及96例健康对照。多因素Logistic回归分析显示,血红蛋白(HGB)计数、血小板(PLT)计数、血小板分布宽度(PDW)、NLR水平是食管鳞状上皮内瘤变的独立预测因子。根据以上四项指标构建的列线图模型曲线下面积(area under curve,AUC)为0.787。Bootstrap内部验证的C-index值为0.771,提示该食管鳞状上皮内瘤变风险预测模型的识别能力、一致性和临床净获益良好。结论基于HGB、PLT、PDW及NLR建立的列线图预测模型可有效鉴别有无食管鳞状上皮内瘤变病变人群,对诊断食管癌前病变有辅助价值。
