The utilization of Mesenchymal stem cells (MSCs)-derived secretome was suggested as a promising alternative in cell-based regenerative therapy. Herein, the MSCs cells were impregnated with a pan-PI3K/Akt/mTOR inhibitor and their secretome was utilized to explore the anticancer and antimetastasis effects against breast cancer. To establish this aim Bone marrow-derived MSCs was treated with 50, 100, or 250 nM Wortmannin (Wort), where the cytotoxic, apoptotic, and autophagic potential of their secretome were investigated in luminal-A breast cancer cells (MCF-7). We found that exposure of MCF-7 to Wort-containing secretome induced both apoptosis and autophagy, whereas prolonged exposure led to massive cell death. Also, Wort-loaded secretome induced nuclear DNA fragmentation and reduced cell metastasis in vitro. These findings were associated with Wort-dependent decrease in the formation of the phosphorylated Akt and mTOR proteins, reduced the expression of their mRNAs, and downregulate of the expression of the catalytic subunit of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K-CA). Taken together, these findings suggested the promising antiproliferative and antimetastasis effects of combining pan-PI3K/Akt/mTOR inhibitors with MSCs-derived secretome in breast cancer.