Background <p>Diabetic foot ulcer (DFU) is chronic complication of diabetes. The aim of this study was to investigate the function and mechanism of lncRNA LINC00641 and miR-362-5p in DFU.</p> Methods <p>The expressions of target genes were detected using RT-qPCR. ROC curve and logistic analysis were used to evaluate the diagnostic ability of LINC00641. High glucose (HG) treatment with 30&#xa0;mM D-glucose of HDFa cells simulates cell damage. The CCK8 assay and flow cytometry were used to detect cell viability and apoptosis. The luciferase reporter assay was used to detect miR-362-5p as the target gene of LINC00641. Pearson correlation analysis was used to analyze the relationship between LINC00641 and miR-362-5p in DFU.</p> Results <p>LncRNA LINC00641 was highly expressed in DFU and showed high diagnostic capacity. HG treatment promoted LINC00641 expression, inhibited cell proliferation, increased apoptosis, elevated MMP1 levels, and decreased VEGF and SDF-1α levels. However, silencing LINC00641 reversed the effects of HG treatment. In addition, LINC00641 has binding sites with miR-362-5p, and the two were negatively correlated. Co-silencing LINC00641 and miR-362-5p reversed the effect of silencing LINC00641 on HDFa cells.</p> Conclusion <p>LncRNA LINC00641 may be used as a diagnostic tool for DFU. Moreover, LINC00641 is involved in DFU healing through the miR-362-5p sponge mechanism.</p>