What question did this study set out to answer?

This study aims to investigate the combined effects of ivermectin and metformin on MCF-7 breast cancer cells, focusing on the underlying mechanisms.

May 18, 2026Oncology Reports0 citations

Combination of ivermectin and metformin promotes autophagy in MCF‑7 cells by inhibiting phosphorylation of the PI3K/AKT/mTOR pathway

HFHuili Feng LHLixin He TUTalha Umar

Key Points

This study aims to investigate the combined effects of ivermectin and metformin on MCF-7 breast cancer cells, focusing on the underlying mechanisms.
Cell Counting Kit‑8 assays assessed proliferative activity of MCF-7 cells.
Transcriptomic analysis identified signaling pathway alterations and differentially expressed genes.
Western blotting and flow cytometry evaluated protein expression and reactive oxygen species levels.
Combined treatment with IVM and MET significantly reduced cell viability and migration in a concentration-dependent manner.
Reduction in phosphorylation of p-PI3K, p-AKT, and p-mTOR levels was observed with combination therapy.
Increased intracellular ROS levels were found in the treatment group, triggering oxidative stress responses and enhancing autophagy.

Abstract

Breast cancer remains one of the leading causes of cancer‑related mortality among women globally, necessitating the exploration of novel therapeutic strategies. The present study investigated the combined effects of Ivermectin (IVM) and metformin (MET) on the human mammary tumor cell line MCF‑7, with a focus on their mechanisms of action. Cell Counting Kit‑8 assays were employed to evaluate proliferative activity, while Transwell migration and scratch assays assessed invasive and migratory capabilities. Transcriptomic analysis identified differentially expressed genes and key signalling pathways, while flow cytometry quantified reactive oxygen species (ROS) levels. Autophagosome formation was visualized using transmission electron microscopy. Western blotting and immunofluorescence staining were employed to detect changes in the expression of key proteins. Results demonstrated that combined IVM and MET intervention significantly inhibited MCF‑7 cell viability after 24 h, showing concentration‑dependent reductions in proliferation, migration and invasiveness. Transcriptomic analysis revealed a significant enrichment of the PI3K/AKT/mTOR signaling pathway, accompanied by decreased expression of thrombospondin‑1 (THBS1). Western blotting revealed that the combination therapy reduced phosphorylation levels of phosphorylated (p‑)PI3K, p‑AKT and p‑mTOR. Through the use of THBS1 overexpression plasmids, the present study validated its positive regulatory role in the PI3K/AKT/mTOR pathway, demonstrating that THBS1 mediates phosphorylation within this signaling cascade. Flow cytometry analysis demonstrated that the IVM combined with MET treatment group exhibited significantly elevated intracellular ROS levels compared with the single‑drug therapy group, triggering oxidative stress responses. After clearing ROS using N‑acetyl‑L‑cysteine, the PI3K/AKT/mTOR signaling pathway was reactivated. Additionally, transmission electron microscopy revealed extensive autophagosome formation within tumor cells of the IVM‑MET combination group. The results indicate that IVM‑MET inhibits PI3K/AKT/mTOR pathway phosphorylation through the accumulation of ROS and the downregulation of THBS1 expression, thereby activating autophagy programs and ultimately leading to tumor cell death.

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