1、J Clin Pathol Res2023,43(6)http:/ 临床与病理杂志FCHSD2：肺鳞状细胞癌潜在的预后生物标志物蒋代顺1，张璐1，刘义21.广东医科大学(湛江校区)药学院，广东 湛江 524000；2.广东医科大学广东天然药物研究与开发实验室，广东 湛江 524000摘要 目的：探讨Fer/Cip4同源性(Fer/Cip4 homology，FCH)和双Src同源性3(Src homology 3，SH3)结构域2(FCH and double SH3 domains 2，FCHSD2)在肺鳞状细胞癌(squamous cell lung cancer，LUSC)中的表达情况，
2、并分析其与患者预后的关系。方法：采用肿瘤免疫评估资源2(Tumor Immune Estimation Resource 2，TIMER2)进行泛癌分析，发现FCHSD2的表达差异。用Kaplan-Meier绘图仪评估FCHSD2对LUSC的预后价值，使用癌症基因组图谱(The Cancer Genome Atlas，TCGA)数据进行确认。并通过定量RT-PCR检测4种NSCLC细胞系和人支气管上皮样细胞中FCHSD2信使RNA(messenger RNA，mRNA)的表达情况。通过Coexpedia肿瘤数局库分析了FCHSD2共表达的前100个基因，对其功能进行富集分析，并在Sangerb
3、ox 3.0平台探究FCHSD2表达与免疫微环境的关系，最终预测 FCHSD2 的调控微 RNA(microRNA，miRNA)，并利用 RNA 相互作用百科全书(Encyclopedia of RNA Interactomes，ENCORI)平台建立LUSC潜在的miRNA-mRNA调控网络。结果：生存分析显示FCHSD2 mRNA水平的降低与LUSC患者OS的降低显著相关。用GO进行功能注释富集分析和KEGG通路富集分析，得到与磷酸转移酶活性、肝素结合等结果相关。且浸润水平随着FCHSD2表达的变化而变化，呈正相关。最终预测出了FCHSD2的调控miRNA，并构建了潜在的miRNA-mRN
4、A调控网络。结论：FCHSD2基因有望成为新型LUSC预后标志物，为LUSC精准治疗策略选择提供重要依据。关键词 肺鳞状细胞癌；预后标志物；FCHSD2FCHSD2:A potential prognostic biomarker for squamous cell lung cancerJIANG Daishun1,ZHANG Lu1,LIU Yi21.School of Pharmacy,Guangdong Medical University(Zhanjiang Campus),Zhanjiang Guangdong 524000;2.Guangdong Natural Medicine
5、 Research and Development Laboratory,Guangdong Medical University,Zhanjiang Guangdong 524000,ChinaABSTRACT Objective:To explore the expression of Fer/Cip4 homology(FCH)and double Src homology 3(SH3)domains 2(FCHSD2)in squamous cell lung cancer(LUSC),and to analyze its DOI：10.11817/j.issn.2095-6959.2
6、023.221970收稿日期(Date of reception)：2022-09-15第一作者(First author)：蒋代顺，Email:,ORCID:0009-0002-7396-8971通信作者(Corresponding author)：刘义，Email:,ORCID:0000-0002-3884-6605基金项目(Foundation item)：国家自然科学基金(82073054)。This work was supported by the National Natural Science Foundation of China(82073054).1074FCHSD2：肺
7、鳞状细胞癌潜在的预后生物标志物 蒋代顺，等relationship with the prognosis of patients.Methods:Tumor Immune Estimation Resource 2(TIMER2)web was used for pan-cancer analysis,and differences in the expression of FCHSD2 were found.The prognostic value of FCHSD2 in LUSC was assessed with a Kaplan-Meier plotter and confirmed
8、 using The Cancer Genome Atlas(TCGA)data.The expression of FCHSD2 messenger RNA(mRNA)in non-small cell lung cancer(NSCLC)cell lines and human bronchial epithelioid cells was detected by quantitative RT-PCR.The top 100 genes co-expressed by FCHSD2 were analyzed through the Coexpedia tumor database.An
9、d their function was analyzed by enrichment analysis,and the relationship between FCHSD2 expression and immune microenvironment were explored on the Sangerbox 3.0 platform.Finally,by predicting the regulatory microRNA(miRNA)of FCHSD2,and using the Encyclopedia of RNA Interactomes(ENCORI)platform,a p
10、otential regulatory network of miRNA mRNA of LUSC was established.Results:Survival analysis showed a significant correlation between a decrease in FCHSD2 mRNA levels and a decrease in OS in LUSC patients.Functional annotation enrichment analysis and KEGG pathway enrichment analysis were conducted us
11、ing GO,and results related to phosphotransferase activity and heparin binding were obtained.And the infiltration level changes with the expression of FCHSD2,showing a positive correlation.Finally,the regulatory miRNA of FCHSD2 was predicted and a potential miRNA mRNA regulatory network was construct
12、ed.Conclusion:FCHSD2 gene is expected to become a new type of LUSC prognostic marker,which provides an important basis for the selection of precise treatment strategies for LUSC.KEY WORDS squamous cell lung cancer;prognostic biomarker;FCHSD2根据2018年全球癌症统计，肺癌在所有肿瘤中发病率和病死率最高。非小细胞肺癌(non-small cell lung
13、cancer，NSCLC)是最常见的病理类型，约占所有肺癌的 85%1-2。在 NSCLC 中，肺鳞状细胞癌(squamous cell lung cancer，LUSC)是第二常见的NSCLC类型，每年有超过 40 万例新发病例，占 NSCLC 的 20%30%。LUSC 患者的 5 年总生存期(overall survival，OS)在临床阶段I和II期约为40%，而在临床阶段III和IV期不到5%3。因此，识别新的预后标志物和治疗靶点对LUSC的临床治疗很重要。目前，许多治疗方法正应用于肺癌，如手术切除、化学疗法、放射治疗、靶向治疗和免疫疗法。早期NSCLC患者常接受外科手术切除肿瘤手术
14、，但对于晚期阶段无法通过手术切除肿瘤的患者，靶向治疗或与化学疗法结合的免疫疗法是最好的治疗方法4-5。本 研 究 拟 探 讨 Fer/Cip4 同 源 性(Fer/Cip4 homology，FCH)和双 Src 同源性 3(Src homology 3，SH3)结 构 域 2(FCH and double SH3 domains 2，FCHSD2)信使RNA(messenger RNA，mRNA)表达与LUSC临床特征之间的联系，以及FCHSD2在LUSC患者预后和免疫调节中的可能作用，并进一步验证FCHSD2在NSCLC细胞系和人支气管上皮样细胞中mRNA的表达情况。1 材料与方法 1.1
15、 TIMER2分析肿瘤免疫评估资源 2(Tumor Immune Estimation Resource 2，TIMER2)6(http:/www.cistrome.org/)是一个癌症微阵列数据库和基于网络的数据挖掘平台，用于分析不同癌症中FCHSD2的转录水平。采用t检验，将FCHSD2在临床肿瘤标本中的mRNA表达水平与正常对照组进行比较。倍数大于1.5，P0.4的交互作用具有统计学意义。Cytoscape9(版本3.7.2)是一个开源生物信息学软件平台，用于可视化分子相互作用网络。1.7 GO功能富集和KEGG通路富集分析基因本体(gene ontology，GO)资源平台10提供基因
16、的功能注释和富集分析。京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes，KEGG)11是一个全面的生物信息数据库，旨在帮助分析大规模的分子数据集。1.8 Sangerbox 3.0免疫浸润分析与免疫细胞分析Sangerbox 3.0(http:/ ENCORI数据库RNA 相互作用百科全书(Encyclopedia of RNA Interactomes，ENCORI；http:/)是一个开源平台，用于研究微RNA(microRNA，miRNA)-mRNA 相互作用。本研究应用 ENCORI 来预测调控FCHSD2的miRNAs，并验证其与RNA表达的相关性。1.10 Kaplan-Meier生存曲线Kaplan-Meier 绘图仪12(https:/ LUSC 中已鉴定的 miRNAs 表达的预后意义，利用数据发现和验证预后生物标志物，基于荟萃分析的在线工具，分析来源多个基因表达和临床数据。为了评估特定miRNA的预后价值，患者样本根据该基因的中位数表达(高与低)将其分为2个队列。本研究获得符合要求的 miRNAs 的 Kapla