COL10A1在胃癌中的作用机制及其临床意义

The Mechanism and Clinical Significance of COL10A1 in Gastric Cancer

摘要: 目的：胃癌(gastric cancer, GC)作为最常见的消化道恶性肿瘤，由于早期症状不明显，发现时多为晚期，预后较差。COL10A1过去主要研究其在骨关节疾病中的作用，而近年来更多地关注于该基因在肿瘤中的作用，其可能通过改变肿瘤微环境的成分，发挥促进肿瘤发生、转移等方面的能力，具有成为多种肿瘤诊断标志物或治疗相关靶点的潜力，而其在胃癌中作用的研究较少，本研究目的在于利用生物信息学分析的方法，探究COL10A1在对于胃癌发生、发展中的作用以及在肿瘤诊断、预后预测及胃癌治疗等方面的临床应用价值。方法：收集TCGA数据库中胃癌患者的相关数据，联合GTEx等其他数据库数据，使用R软件中相关R包分析COL10A1在胃癌与普通组织中的表达差异。进而分析COL10A1高低表达组之间的基因表达情况得到差异表达基因，对这些差异表达基因进行功能富集分析(GO, KEGG, GSEA)，探究目标基因COL10A1差异表达可能影响的细胞功能、通路。使用Estimate算法对COL10A1高低表达组样本进行打分，然后比较高低表达组之间评分，评估COL10A1的表达差异对细胞外基质的影响。同时使用CIBERSORT算法对COL10A1高低表达组中样本的免疫细胞浸润进行分析，比较高低表达组之间免疫细胞浸润情况的不同，分析COL10A1对于肿瘤组织中的免疫细胞浸润的影响。为探究COL10A1在临床中的作用，首先根据TCGA数据库中临床数据进行生存分析，分析COL10A1对于总生存期、无病生存期的影响，进而对COL10A1表达量及其他常见因素(如年龄、性别、TNM分期、Stage分期等)进行单因素COX分析与多因素COX分析，并构建多因素COX风险比例模型。根据TCIA数据库中数据比较目标基因高低表达组之间免疫治疗效果，并评估COL10A1与常见免疫检查点表达之间表达相关性，探究其与免疫治疗的相关性。通过上述方法探究COL10A1在胃癌中可能参与的机制、对于预后、免疫治疗效果的影响。结果：通过分析TCGA、GTEx数据库数据，目标基因COL10A1在包括胃癌在内的多种肿瘤中显著高表达，结合所收集样本测序结果，目标基因在胃癌组织与正常胃组织中表达差异明显。通过分析数据库中数据表明COL10A1表达量与患者预后相关，对于总生存期，高表达胃癌患者的总生存期较短，预后较差。经单因素及多因素COX分析，COL10A1表达量是患者预后的独立危险因素，并根据TCGA数据库数据，构建了预后预测模型。在细胞外基质的组成方面发挥作用，同时可能参与了TGF-β通路及PI3K-Akt信号通路，两种通路均与肿瘤的发生发展相关。通过进行Estimated、CIBERSORT分析，COL10A1高低表达组患者细胞外基质差异明显，高表达组患者肿瘤纯度更低、免疫细胞浸润数量更多，于是其表达与肿瘤微环境中免疫细胞浸润呈正相关。高表达组免疫治疗效果较差，可能与目标基因和常见免疫检查点表达呈正相关有关。预示着该基因有着作为治疗靶点的潜力。结论：COL10A1在在胃癌中高表达，当COL10A1表达上调时胃癌患者的总体生存期显著缩短，但对无进展生存期无明显影响，提示其与患者的预后不良有关。COL10A1与肿瘤微环境的构成有关，与免疫细胞浸润和免疫检查点的表达存在显著性相关。通过分析TCIA数据库中数据，COL10A1高表达会使部分患者的免疫治疗效果差，提示COL10A1可能是一个潜在的免疫治疗靶点。

Abstract: Objective: Gastric cancer (GC), as the most common malignant tumor of the digestive tract, is often found in the late stage due to its unclear early symptoms and poor prognosis. COL10A1 used to mainly study its role in bone and joint diseases, but in recent years more attention has been paid to the role of this gene in tumors. It may play a role in promoting tumor occurrence, metastasis and other aspects by changing the composition of the tumor microenvironment, and has the potential to become a variety of tumor diagnostic markers or treatment related targets, while there is less re-search on its role in gastric cancer. The purpose of this study is to use bioinformatics analysis methods to explore the role of COL10A1 in the occurrence and development of gastric cancer, as well as its clinical application value in tumor diagnosis, prognosis prediction, and treatment of gas-tric cancer. Method: Collect relevant data of gastric cancer patients in the TCGA database, combined with data from other databases such as GTEx, and analyze the expression difference of COL10A1 between gastric cancer and common tissues using the relevant R package in R software. Further analysis of the gene expression between the high and low expression groups of COL10A1 was con-ducted to identify differentially expressed genes. Functional enrichment analysis (GO, KEGG, GSEA) was performed on these differentially expressed genes to explore the potential cellular functions and pathways affected by the differential expression of the target gene COL10A1; use the Estimate algorithm to score the samples of COL10A1 high and low expression groups, and then compare the scores between the high and low expression groups to evaluate the impact of COL10A1 expression differences on the extracellular matrix. Simultaneously, the CIBERSORT algorithm was used to ana-lyze the immune cell infiltration of samples in the high and low expression groups of COL10A1, compare the differences in immune cell infiltration between the high and low expression groups, and analyze the impact of COL10A1 on immune cell infiltration in tumor tissue. To explore the role of COL10A1 in clinical practice, survival analysis was first conducted based on clinical data from the TCGA database, analyzing the impact of COL10A1 on overall survival and disease-free survival. Then, single factor COX analysis and multi factor COX analysis were conducted on the expression level of COL10A1 and other common factors (such as age, gender, TNM stage, Stage stage, etc.), and a multi factor COX risk proportion model was constructed. To analyze the expression correlation between COL10A1 and common immune checkpoint expression, to delve into its relevance to immunother-apy, and to compare the effect of immunotherapy between groups of high and low expression of target genes using data from the TCIA database, the drug sensitivity of high and low expression groups to some chemotherapeutic agents was analyzed using pRRophetic. Through the above methods, we investigated the possible mechanisms of COL10A1 involvement in GC and its impact on the prognosis and immunotherapeutic effect. Result: By analyzing the TCGA and GTEx database da-ta, the target gene COL10A1 was significantly over expressed in various tumors, including gastric cancer. Combined with the sequencing results of the collected samples, the target gene showed sig-nificant differences in expression between gastric cancer tissue and normal gastric tissue. By ana-lyzing the data in the database, it was found that the expression level of COL10A1 is related to pa-tient prognosis. For overall survival, high expression gastric cancer patients have a shorter overall survival period and a poorer prognosis. After univariate and multivariate COX analysis, the expres-sion of COL10A1 was found to be an independent risk factor for patient prognosis. Based on the TCGA database data, a prognosis prediction model was constructed. Analysis by GO, KEGG, and GSEA concluded that COL10A1 plays a role in the composition of the extracellular matrix and may be in-volved in the TGF-β pathway and PI3K-Akt signaling pathway, both of which are usually risk factors for tumor development according to the current findings. Through the estimated and CIBERSORT analysis, the extracellular matrix of patients with high and low expression of COL10A1 was signifi-cantly different. The tumor purity of patients with high expression of COL10A1 was lower, and the number of immune cells infiltrated was more. Therefore, its expression was positively correlated with the infiltration of immune cells in the tumor microenvironment. The immunotherapy effect of the high expression group is poor, which may be positively correlated with the expression of target genes and common immune checkpoints, indicating the potential of this gene as a therapeutic tar-get. Conclusion: COL10A1 is highly expressed in gastric cancer. When COL10A1 expression is up-regulated, the overall survival period of gastric cancer patients is significantly shortened, but it has no significant impact on progression free survival, indicating that it is related to poor prognosis of patients. COL10A1 is related to the composition of tumor microenvironment, and is significantly re-lated to the infiltration of immune cells and the expression of immune checkpoints. By analyzing the data in the TCIA database, it was found that high expression of COL10A1 can lead to poor im-munotherapy efficacy in some patients, suggesting that COL10A1 may be a potential immunother-apy target.