Dermatan sulfate (DS) is a glycosaminoglycan (GAG) that is produced through the epimerization of the glucuronic acid on chondroitin sulfate into iduronic acid (IduA) by dermatan sulfate epimerase (DS-epi) 1 and 2. Proteoglycans (PGs) play essential physiological and pathological roles during cellular development, proliferation, differentiation, and cancer metastasis. DS proteoglycans play vital roles during the process of tumorigenesis, due to the increased flexibility of the polysaccharide chain in the presence of IduA residues, which facilitate specific interactions with proteins, such as growth factors, cytokines, and angiogenic factors. Furthermore, DS-epi is highly expressed in many tumors, especially in esophageal squamous cell carcinoma. This study aimed to investigate the expression of DS-epi1 in multiple breast cancer cell lines, including MCF7 (luminal A), MDA-MB-231 (triple-negative) and SKBR3 (human epidermal growth factor receptor 2-positive), and its involvement in cancer progression. A SKBR3 variant, SKBR3m, presented the most erratic cell growth pattern when compared with those for MCF7 and MDA-MB-231. Moreover, SKBR3m cells demonstrated the highest level of DS-epi1 gene expression and higher S-DS content. However, at the protein level, MCF7 cells displayed the highest protein level for DS-epi1, whereas MDA-MB-231 cells had the lowest level. DS-epi1 was found in vesicles and in the perinuclear compartment only in SKBR3m cells, suggesting localization in the Golgi apparatus in these cells, in contrast with the cytoplasmic localization observed in MCF7 and MDA-MB-231 cells. The cytoplasm location of DS-epi1 likely compromised the formation of DS chains, but the core protein was detected using a decorin antibody. Golgi-specific labeling confirmed the localization of DS-epi1 in SKBR3m cells at the Golgi apparatus, indicating that the location of the enzyme was a determinant for the synthesis of DS in this cell line, suggesting that DS may play a decisive role in the tumor growth observed in this breast cancer cell line.Copyright © 2020 Elsevier Ltd. All rights reserved.