基于生物信息学方法分析LAYN在甲状腺癌中临床预后价值及对免疫细胞浸润的影响

目的 探索LAYN基因在甲状腺癌组织中的表达与临床预后的相关性及对免疫细胞浸润的影响,为进一步研究潜在的分子机制提供基础。方法 在Oncomine和GEPIA数据库中检索甲状腺癌相关的基因,分析LAYN基因在甲状腺癌组织与正常组织之间表达的差异。通过GEPIA数据库分析LAYN基因与甲状腺癌患者临床分期及预后的相关性。结合ImmucellAI及TIMER2.0工具分析LAYN基因与甲状腺癌组织中CD4⁺T细胞和CD8⁺T细胞浸润丰度的关系。联合STRING数据库分析LAYN表达相关的蛋白。通过实时荧光定量PCR(RT-PCR)进一步验证检测甲状腺癌组织及癌旁组织,甲状腺癌细胞系SW579,TPC-1及甲状腺上皮细胞TEC中LAYN的表达水平。结果 Oncomine、GEPIA和STRING数据库结果表明,与正常甲状腺组织相比,LAYN基因在甲状腺癌组织中低表达(P＜0.05)。LAYN基因与甲状腺癌临床TNM分期相关(P＜0.05),Ⅲ/Ⅳ期患者肿瘤组织中LAYN基因低表达。LAYN基因高表达组患者总生存期(Overall survival,OS)较差,而无疾病生存期(Disease free survival,DFS)较好(P＜0.05)。在甲状腺癌组织中,CD4⁺T细胞和CD8⁺T细胞的浸润丰度较低,且CD4⁺T细胞和CD8⁺T细胞的浸润丰度与LAYN基因的表达水平呈正相关(P＜0.05)。此外,LAYN相互作用的蛋白为PNRC1,MAGEH1,HAPLN1,SMOC1,PRSS12,HMMR,NF2,TLN2,TLN1,TLE2。RT-qPCR验证结果表明,LAYN在甲状腺癌及甲状腺癌细胞系SW579及TPC-1中低表达(P＜0.05)。结论 LAYN基因在甲状腺癌组织中低表达,且与甲状腺癌患者临床TNM分期及预后相关,可作为预后判断的分子标志物。其潜在机制可能通过影响浸润性免疫细胞丰度发挥作用。为研究甲状腺癌致病机制及靶向药物的开发提供理论依据。

Analysis for clinical prognostic value of LAYN in thyroid cancer and its influence on infiltration of immune cells

Objective The objective of this study was to explore the correlation between the expression of LAYN gene in thyroid cancer tissues and clinical prognosis,and its influence on infiltration of immune cell,so as to provide a basis for further research on potential molecular mechanism. Methods The genes related to thyroid cancer were collected from Oncomine and GEPIA databases,and analyzed the differences of the expression of LAYN gene between thyroid cancer tissues and normal tissues.The GEPIA database was used to analyze the correlation between the LAYN gene and the clinical stage,and prognosis of patients with thyroid cancer.The combination of ImmucellAI and TIMER2.0 tools was used to analyze the relationship between the LAYN gene and CD4⁺T cells and CD8⁺T cells infiltration abundance in thyroid cancer tissues.Combination with the STRING database was used to analyze the LAYN expression related proteins.Real-time fluorescent quantitative PCR(RT-qPCR)was used to further verify and detect the expression of LAYN in thyroid cancer tissues and adjacent tissues,thyroid cancer cell lines SW579,TPC-1 and thyroid epithelial TEC cells. Results The results of Oncomine,GEPIA and STRING database showed that compared with normal thyroid tissues,LAYN gene was low expressed in thyroid cancer tissues(P＜0.05).The LAYN gene was related to the clinical TNM staging of thyroid cancer(P＜0.05),and the LAYN gene was low expressed in tumor issues of patients with stage Ⅲ/Ⅳ.The overall survival(OS)of patients in the high expression of LAYN gene group was poor,while the disease free survival(DFS)was better(P＜0.05).In thyroid cancer tissues,the infiltration abundance of CD4⁺T cells and CD8⁺T cells was low,and the infiltration abundance of CD4⁺T cells and CD8⁺T cells was positive correlation with the expression of LAYN gene(P＜0.05).In addition,the proteins related to the expression of LAYN were PNRC1,MAGEH1,HAPLN1,SMOC1,PRSS12,HMMR,NF2,TLN2,TLN1 and TLE2.The verification results of RT-PCR showed that LAYN was low expressed in thyroid cancer and thyroid cancer SW579 and TPC-1 cell lines(P＜0.05). Conclusion The LAYN gene is low expressed in thyroid cancer tissues,and it is related to the clinical TNM staging and prognosis of patients with thyroid cancer.It can be used as a molecular marker for prognostic judgment.The underlying mechanism of this gene may play a role in affecting the abundance of infiltrating immune cells.This study provides a theoretical basis for studying the pathogenesis of thyroid cancer and the development of targeted drugs.