Inhibition of MID1IP1 induces ferroptosis and suppresses c-Myc expression in colorectal cancer cell

Background: Colorectal Cancer (CRC) is the third most diagnosed Cancer worldwide. Despite treatment advances, CRC still requires novel strategies to overcome drug resistance and metastasis. Midline 1 Interacting Protein 1 (MID1IP1) is highly expressed in cancers and promotes tumor growth through c-Myc-mediated ribosomal protein regulation. We hypothesized that MID1IP1 contributes to CRC progression via c-Myc and Ferroptosis.

Objective: To investigate how MID1IP1 affects metabolic reprogramming, Ferroptosis, and migration in CRC.

Methods: MID1IP1 was silenced in HCT116 cells using siRNA. Colony formation assays evaluated growth. Western blotting assessed c-Myc, ferroptosis-related proteins, glycolytic Enzymes, and EMT markers. Intracellular ROS was measured with a fluorescence probe, and a Ferroptosis inhibitor confirmed ferroptotic cell death. Immunofluorescence detected c-Myc and PKM2, while wound healing assays evaluated migration.

Results: MID1IP1 knockdown reduced cell growth and c-Myc stability, lowering glycolysis-related protein expression. GPX4 and Other ferroptosis-protective proteins decreased, resulting in ROS accumulation and Ferroptosis. Migration was inhibited, with altered EMT marker expression.

Conclusion: MID1IP1 promotes CRC cell survival by stabilizing c-Myc and preventing Ferroptosis, while also influencing migration. These findings suggest that MID1IP1 may serve as a novel biomarker and therapeutic target in CRC.