Objective MSC-derived exosomes (MSC-Exos) from peripheral blood Mesenchymal stem cells (MSCs) were studied for their influence on epithelial-mesenchymal stem cells (MSCs) from A549 cells with non-small cell lung cancer (NSCLC). Methods Oil red O was used to stain MSCs isolated from peripheral blood of NSCLC patients to determine their lipid differentiation ability. An ultrafast centrifugation method was used to separate MSCs and exosomes, transmission electron microscopy was used to view exosome morphology, and NTA particle size analysis was used to describe their size and distribution. CD9, TSG101, and calnexin were identified in MSC-Exos through Western blot analysis, while miR-494-3p expression was quantified using RT-qPCR. In order to evaluate the influence of MSC-Exos on the migratory and invasive capabilities of NSCLC cells, A549 cells were cultivated and subsequently subjected to a Transwell cell invasion assay. In addition to RT-qPCR analysis of ZEB1 mRNA expression, luciferase assays were performed to determine miR-494-3p binding to ZEB1. Immunofluorescence staining was utilized for the evaluation of E-cadherin and Vimentin expression levels, and Western blot analysis was employed to determine the levels of ZEB1, E-cadherin, Vimentin, MMP2, and MMP9. Results MiR-494-3p expression in MSCs-Exos was significantly reduced, inhibiting the proliferation, migration, and EMT processes in A549 cells. As miR-494-3p directly targets ZEB1, it was attenuated when ZEB1 was overexpressed in A549 cells, preventing miR-494-3p from inhibiting EMT.Conclusion As a result of miR-494-3p expression in peripheral blood MSCs from NSCLC patients, A549 cells were inhibited in their EMT process by targeting ZEB1