Objective: Colorectal cancer (CRC) is the third most prevalent malignant tumor type and the second leading cause of cancer-related death. Sequence similarity family 50 member A (FAM50A) plays a vital role in numerous disease processes, including tumor progression. This study aimed to evaluate the prognostic significance of FAM50A in CRC and to explore its role in CRC cell proliferation. Methods: The Cancer Genome Atlas (TCGA) database was used to assess the mRNA expression levels of FAM50A in CRC. Immunohistochemical staining was used to study the expression of FAM50A in CRC tissues. Patient survival data were used to assess the prognostic significance of FAM50A in CRC using Kaplan–Meier analysis and Cox regression analysis. The Cell Counting Kit-8, 5-ethynyl-2′-deoxyuridine, and colony-formation assays were employed to assess the impact of FAM50A knockdown on tumor cell proliferation. Co-expression and correlation heatmaps were used to identify genes whose expression levels were highly correlated with FAM50A, and to further investigate the possible mechanism of action of FAM50A in CRC cell proliferation. Results: Analysis of data from TCGA database revealed a notable upregulation of FAM50A expression levels in CRC tissue compared with adjacent normal tissue. Moreover, FAM50A expression was positively correlated with N, M, and TNM stages in 145 patients with CRC. Cox regression analysis and construction of a nomogram revealed that high FAM50A expression was a prognostic indicator for poor overall survival in patients with CRC. Knockdown of FAM50A decreased cell proliferation ability, the proportion of EdU positive cells, and the number of CRC cell colonies, whereas overexpressing FAM50A promoted proliferative phenotypes. The expression levels of three genes in CRC were significantly negatively correlated, while the expression levels of 18 genes were significantly positively correlated, with the expression level of FAM50A. Conclusion: FAM50A may be a novel prognostic marker for CRC, and may participate in regulating tumor progression by interacting with some highly correlated molecules that drive CRC cell proliferation.