Background: Despite recent advances in colorectal cancer (CRC) diagnosis and population screening programs, the identification of patients with preneoplastic lesions or with early CRC stages remains challenging and is essential for reducing CRC incidence and increasing patients' survival. Methods: Our study comprehensively investigated the clinical significance and relationship among AGER complex protein/N-Glycosilation genes (DDOST, PRKCSH, and GALECTIN 3) and colon cancer progression in colorectal cancer through a bioinformatics data mining process (STRING, UALCAN, HPA, GEPIA2, TNMplot), their interaction (DMFold-Multimer), and followed by experimental validation. We analyzed 112 colorectal tissue samples originating from CRC stages I to IV, including normal tissue and adenomas. The characterization of three biomarker candidates was performed using immunohistochemistry on normal tissue, precancerous, and cancerous lesions with increasing CRC stages. We selected the ROI in QuPath and analysed it in ImageJ using the DeepLIIF extension to calculate expression percentage. Results: The distributions of PRKCSH, DDOST, and GALECTIN 3 were validated in tissues, showing different expression levels, especially in early stages of CRC, compared to normal and preneoplastic tissues. Conclusion: We highlighted three proteins that require further investigation to better characterize their role in early CRC carcinogenesis and their potential as markers of CRC progression.