<b>Systematic Analysis and Mechanistic Exploration of Glyoxalase I Inhibitor Modulation in Breast Cancer-Associated Endothelial Cells: An Integrated Approach of Network Pharmacology, Bioinformatics, and Experimental Validation</b>

Breast cancer angiogenesis plays a critical role in the progression of breast cancer. Glyoxalase I (GLO1), while primarily studied for its role in endothelial cells within cardiovascular and metabolic diseases, has an unclear role in breast cancer-associated endothelial cells (BCECs). Additionally, a systematic review of GLO1 inhibitors is lacking. This study aims to systematically analyze the pharmacological profiles of GLO1 inhibitors, screen for those that regulate BCECs, and validate these findings experimentally. The pharmacological profiles of 17 GLO1 inhibitors were analyzed using SwissADME and the pkCSM tool, while target identification was performed using SwissTargetPrediction and SuperPred. A network of potential targets was constructed by intersecting these with differentially expressed genes (DEGs) from the GSE80506 dataset of BCECs. This network was subjected to enrichment analysis, expression correlation analysis, and molecular docking. The results were then validated through cellular assays and aortic ring assay in mice. A total of 515 targets were associated with the 17 GLO1 inhibitors, of which 36 were identified as potential targets related to BCEC regulation. Enrichment analysis revealed that the cell cycle is a critical pathway regulated by GLO1 inhibitors, with CCNA2, CCNE1, CDC25A, CDC25B, CDK6, CHEK1, PLK1, and TTK being key targets. Molecular docking indicated that <i>Glyoxalase I inhibitor 6</i> plays a significant role in regulating these targets. Cellular assays and aortic ring assay in mice demonstrated that this inhibitor blocked cells in the G0/G1 phase by modulating CCNA2 and CCNE1, and effectively inhibited proliferation, migration, and angiogenesis in BCECs, while also promoting cell death. GLO1 inhibitors show potential regulatory effects on BCECs. Specifically, <i>Glyoxalase I inhibitor 6</i> inhibited proliferation, cell cycle progression, migration, and angiogenesis in these cells. These findings suggest a novel approach for utilizing GLO1 inhibitors in targeting tumor-associated endothelial cells.