Data_Sheet_1_Development and Clinical Validation of a Novel 4-Gene Prognostic Signature Predicting Survival in Colorectal Cancer.zip

In this study, we collected genes related to energy metabolism, used gene expression data from public databases to classify molecular subtypes of colon cancer (COAD) based on the genes related to energy metabolism, and further evaluated the relationships between the molecular subtypes and prognosis and clinical characteristics. Differential expression analysis of the molecular subtypes yielded 1948 differentially expressed genes (DEGs), whose functions were closely related to the occurrence and development of cancer. Based on the DEGs, we constructed a 4-gene prognostic risk model and identified the high expression of FOXD4, ENPEP, HOXC6, and ALOX15B as a risk factor associated with a high risk of developing COAD. The 4-gene signature has strong robustness and a stable predictive performance in datasets from different platforms not only in patients with early COAD but also in all patients with colon cancer. The enriched pathways of the 4-gene signature in the high- and low-risk groups obtained by GSEA were significantly related to the occurrence and development of colon cancer. Moreover, the results of qPCR, immunohistochemistry staining and Western blot assay revealed that FOXD4, ENPEP, HOXC6, and ALOX15B are over expressed in CRC tissues and cells. These results suggesting that the signature could potentially be used as a prognostic marker for clinical diagnosis.

本研究收集了能量代谢相关基因，依托公共数据库中的基因表达数据，基于上述能量代谢相关基因对结肠癌（COAD）开展分子亚型分类，并进一步评估了该分子亚型与患者预后及临床特征之间的关联。对各分子亚型进行差异表达分析后，共得到1948个差异表达基因（DEGs），其功能与癌症的发生发展密切相关。基于该批差异表达基因，我们构建了一款四基因预后风险模型，并确定FOXD4、ENPEP、HOXC6及ALOX15B的高表达可作为与COAD高发病风险相关的风险因子。该四基因特征具备优异的稳健性，在不同平台的数据集、早期COAD患者队列乃至全部结直肠癌患者队列中，均表现出稳定的预测效能。通过基因集富集分析（GSEA）对高低风险组进行富集通路分析发现，该四基因特征富集的通路均与结直肠癌的发生发展显著相关。此外，实时定量PCR、免疫组化染色及蛋白质印迹实验结果显示，FOXD4、ENPEP、HOXC6及ALOX15B在结直肠癌（CRC）组织与细胞中均呈过表达状态。上述结果表明，该基因特征有望作为临床诊断的潜在预后标志物。