目的 以生物信息学方法研究热休克蛋白A4(HSPA4)在肺腺癌中的表达及其临床意义。方法 下载TCGA和GEO数据资料,R软件分析HSPA4基因在肺腺癌和癌旁组织中的表达差异和临床分期的相关性;以HPA数据库资料比较肺腺癌与正常肺组织蛋白表达差异;通过ROC曲线、时间依赖性ROC曲线、列线图、相关性分析HSPA4表达在肺腺癌中的诊断、预后价值和肿瘤微环境中与各种免疫细胞的相关性。应用STRING数据库建立HSPA4相关蛋白互作网络,并利用GO、KEGG及GSEA数据库分析HSPA4及其互作蛋白在肺腺癌中的潜在功能。结果 在TCGA-LUAD、GSE43448和GSE10072数据集中,HSPA4在肺腺癌中mRNA较正常肺组织表达升高(P＜0.001),在蛋白水平表达也明显升高。HSPA4高表达与肺腺癌的TNM分期和病理分期相关;ROC曲线显示HSPA4具有良好的诊断价值(AUC=0.810)。时间依赖性ROC曲线显示1、3、5年AUC分别为0.605、0.583和0.553。列线图显示HSPA4与其他临床因素共同具有较好的1、3、5年生存率预测价值。K-M图显示HSPA4高表达与肺腺癌患者的不良预后有关。HSPA4基因改变率为2.2%,3个甲基化位点与肺腺癌的预后相关。在肿瘤微环境中,HSPA4表达与辅助T细胞、辅助T细胞2等免疫细胞正相关;与B细胞、浆细胞样树突状细胞、CD8 T细胞、细胞毒性细胞、CD56较高表达自然杀伤细胞、T 细胞等免疫细胞呈负相关。PPI蛋白互作显示HSPA4与HSP90AA1、SNCA等蛋白相互作用,GO、KEGG、GSEA通路富集分析显示HSPA4具有参与抗原加工提呈、调节免疫等功能。结论 HSPA4在肺腺癌中高表达,且其表达水平与肺腺癌患者总体预后相关,同时参与了肿瘤的发生和发展,并影响了肿瘤微环境,可能成为肺腺癌中新的生物标记物和治疗靶点之一。
Abstract
Objective To study the expression and clinical significance of heat shock protein A4 (HSPA4) in lung adenocarcinoma by bioinformatics.Methods The data of TCGA and GEO were downloaded, and the expression difference of HSPA4 gene in lung adenocarcinoma and adjacent tissues was analyzed by R software; the differences of protein expression between lung adenocarcinoma and normal lung tissue were compared with HPA database; the diagnostic and prognostic value of HSPA4 expression in lung adenocarcinoma and the correlation between HSPA4 expression and various immune cells in tumor microenvironment were analyzed by ROC curve, time-dependent ROC curve, nomogram and correlation analysis. The STRING database was used to establish HSPA4 related protein interaction network, and GO, KEGG and GSEA databases were used to analyze the potential functions of HSPA4 and its interaction proteins in lung adenocarcinoma.Results In the data sets of TCGA-LUAD, GSE43448 and GSE10072, the expression of HSPA4 in lung adenocarcinoma was higher than that in normal lung tissues and adjacent tissues (P<0.001), and the expression of HSPA4 in lung adenocarcinoma was significantly higher. High expression of HSPA4 was associated with TNM and pathological stages of lung adenocarcinoma; ROC diagram showed that HSPA4 had good diagnostic value (AUC=0.810). The time dependent ROC chart showed that the AUC of 1 year, 3 years and 5 years were 0.605, 0.583 and 0.553 respectively. The nomogram showed that HSPA4, together with other clinical factors, had a good predictive value for 1-year, 3-year and 5-year survival rates. K-M diagram showed that the high expression of HSPA4 was related with the poor prognosis of lung adenocarcinoma patients. The change rate of HSPA4 gene was 2.2%. Three methylation sites were associated with the prognosis of lung adenocarcinoma. In TME, HSPA4 expression was positively correlated with Th2, T helper cells and other immune cells; It was negatively correlated with B cells, pDC, CD8 T cells, Cytotoxic cells, NK CD56bright cells, T cells and other immune cells. PPI protein interaction showed that HSPA4 interacted with HSP90AA1, SNCA and other proteins. Enrichment analysis of GO, KEGG, GSEA pathways showed that HSPA4 had the functions of antigen processing and presenting, and regulating immunity.Conclusions HSPA4 is highly expressed in lung adenocarcinoma, and its expression level is related with the overall prognosis of lung adenocarcinoma patients. It also participates in the occurrence and development of tumor, and affects the tumor microenvironment, which may become a new biomarker and therapeutic target in lung adenocarcinoma.
关键词
热休克蛋白 /
肺腺癌 /
热休克蛋白A4 /
生物信息学
Key words
heat shock protein /
lung adenocarcinoma /
HSPA4 /
bioinformatics
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Siegel R L, Miller K D, Fuchs H E, et al. Cancer statistics, 2022[J]. CA Cancer J Clin, 2022,72(1): 7-33.
[2] Duma N, Santana-Davila R, Molina J R, et al. Non-small cell lung cancer: epidemiology, screening, diagnosis, and treatment [J]. Mayo Clin Proc, 2019, 94(8): 1623-1640.
[3] Chen Z, Fillmore C M, Hammerman P S, et al. Non-small-cell lung cancers: a heterogeneous set of diseases [J]. Nat Rev Cancer, 2014, 14(8): 535-546.
[4] Yang Z, Zhuang L, Szatmary P, et al. Upregulation of heat shock proteins (HSPA12A, HSP90B1, HSPA4, HSPA5 and HSPA6) in tumour tissues is associated with poor outcomes from HBV-related early-stage hepatocellular carcinoma [J]. Int J Med Sci, 2015, 12(3): 256-263.
[5] Jo Y S, Choi M R, Song S Y, et al. Frameshift mutations of HSPA4 and MED13 in gastric and colorectal cancers[J]. Pathol Oncol Res,2016,22(4): 769-772.
[6] Wang Y, Zhao Q, Lan N, et al. Identification of methylated genes and miRNA signatures in nasopharyngeal carcinoma by bioinformatics analysis[J]. Mol Med Rep,2018 , 17(4): 4909-4916.
[7] Ma C, Xu T, Sun X, et al. Network pharmacology and bioinformatics approach reveals the therapeutic mechanism of action of baicalein in hepatocellular carcinoma [J]. Evid Based Complement Alternat Med,2019, 2019:7518374.
[8] Wu C Y, Lin C T, Wu M Z, et al. Induction of HSPA4 and HSPA14 by NBS1 overexpression contributes to NBS1-induced in vitro metastatic and transformation activity [J]. J Biomed Sci,2011,18(1): 1.
[9] Zou T, Liu J Y, She L, et al. The association between heat-shock protein polymorphisms and prognosis in lung cancer patients treated with platinum-based chemotherapy[J]. Front Pharmacol,2020, 11:1029.
[10] Uhlén M, Fagerberg L, Hallström B M, et al. Proteomics. Tissue-based map of the human proteome [J]. Science, 2015,347 (6220): 1260419.
[11] Cerami E, Gao J, Dogrusoz U, et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data [J]. Cancer Discov, 2012, 2 (5): 401-404.
[12] Modhukur V, Iljasenko T, Metsalu T, et al. MethSurv: a web tool to perform multivariable survival analysis using DNA methylation data [J]. Epigenomics, 2018, 10 (3): 277-288.
[13] Szklarczyk D, Gable A L, Lyon D, et al. String v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets [J]. Nucleic Acids Res, 2019, 47 (D1): D607-D613.
[14] Saleh A, Srinivasula S M, Balkir L, et al. Negative regulation of the Apaf-1 apoptosome by Hsp70[J]. Nat Cell Biol,2000,2(8): 476-483.
[15] Dunbar C E, High K A, Joung J K, et al. Gene therapy comes of age[J]. Science,2018,359(6372): eaan4672.
[16] Borst J, Ahrends T, Bąbała N, et al. CD4(+) T cell help in cancer immunology and immunotherapy [J]. Nat Rev Immunol, 2018, 18 (10): 635-647.
[17] Wang Z, Chimenti M S, Strouse C, et al. T cells, particularly activated CD4(+) cells, maintain anti-CD20-mediated NK cell viability and antibody dependent cellular cytotoxicity [J]. Cancer Immunol Immunother, 2022,71 (2): 237-249.
[18] Leong T L, Bryant V L. B cells in lung cancer-not just a bystander cell: a literature review [J]. Transl Lung Cancer Res, 2021, 10 (6): 2830-2841.
[19] Sholl L M. Biomarkers of response to checkpoint inhibitors beyond PD-L1 in lung cancer [J]. Mod Pathol,2022,35(Suppl 1): 66-74.
[20] Doroshow D B, Sanmamed M F, Hastings K, et al. Immunotherapy in non-small cell lung cancer: facts and hopes [J]. Clin Cancer Res, 2019, 25 (15): 4592-4602.
PDF(2475 KB)
