Background: Colorectal cancer, an invasive tumor originating in the mucosal lining of the large intestine and rectum, represents a prevalent form of gastrointestinal malignancy. Although extensive investigations have been conducted on colorectal cancer, the precise molecular mechanisms underlying this neoplasm remain uncharacterized. Alterations in the genetic material have been linked to colorectal cancer progression. To analyze the prognosis of individuals with colorectal cancer, it is crucial to identify new biomarkers. Using integrated bioinformatics analysis, this study successfully identified and confirmed the fundamental gene linked to colorectal cancer. Methods: Bioinformatics software tools were employed to assess the mRNA expression level of Erythroferrone (ERFE) by analyzing the Cancer Genome Atlas (TCGA) dataset, which included 647 tumor samples and 51 control samples. In order to verify the findings, a comparison was conducted with data available on the Gene Expression Omnibus (GEO). In order to determine the clinical significance and expression level of ERFE, a bioinformatics investigation was carried out using logistic regression analysis. To assess survival rates in both high- and low-expression ERFE groups, univariate and multivariate Cox proportional hazards model (Cox) regression analyses were conducted. To validate the expression of ERFE at both the gene and protein levels in colorectal cancer (CRC) cells (HCT116 and Lovo) and normal epithelial cells, reverse transcription quantitative polymerase chain reaction and western blot experiments were performed. The knock-down efficiency of ERFE in HCT116 and LoVo cell lines was assessed using western blot experiments. The effect of ERFE gene function was compared and analyzed through various assays such as cell counting kit-8 (CCK-8), Transwell, and scratch tests before and after ERFE knock-down in HCT116 and LoVo cell lines. Results: An analysis of data obtained from TCGA and GEO databases unveiled a remarkable increase in the expression of ERFE mRNA in CRC tissue compared to both normal and paracancerous tissues (p < 0.001). Furthermore, it was discovered that the overexpression of ERFE was linked to an unfavorable prognosis and had the potential to act as an independent prognostic indicator for predicting overall survival (OS), progression-free interval (PFI), and disease-specific survival (DSS) among colorectal cancer patients. Additionally, a positive correlation was found between heightened ERFE expression, infiltration of numerous immune cells, and levels of immune checkpoint molecules. Experimental evaluations, including the CCK-8 assay, scratch assay, and transwell assay, provided compelling evidence demonstrating a significant reduction in the proliferative, migratory, and invasive capabilities of colorectal cancer cells upon knocking down ERFE. Conclusions: In summary, ERFE, functioning as a pro-oncogene in CRC, is linked to the initiation and advancement of cancer, and can serve as a standalone marker for unfavorable prognosis among CRC patients.