Development of a mitochondrial fluorescent probe reveals anti-colorectal cancer activity via the ZNF124/YES1/YAP1 axis

Purpose Mitochondria are acknowledged to play diverse and critical roles in the development of malignant tumors. Therapeutic agents targeting mitochondria present promising opportunities for cancer treatment. This study aims to synthesize a mitochondrial fluorescent probe, designated MTC-05, and to assess its antitumor efficacy against colorectal cancer (CRC) both in vitro and in vivo . Methods The subcellular localization of MTC-05 was investigated using Mito-Tracker Green FM staining. The antitumor effects of MTC-05 on colorectal cancer cells were evaluated in vitro through assays including cell cycle analysis, colony formation, EdU incorporation, apoptosis detection, wound healing, and trans-well migration assays. Additionally, a xenograft mouse model was employed to assess the therapeutic impact of MTC-05 in vivo . RNA sequencing and α-ketoglutarate level measurements were conducted to elucidate the underlying molecular mechanisms. Results The findings demonstrated that MTC-05 selectively localizes to mitochondria and effectively inhibits malignant phenotypes of CRC. Specifically, MTC-05 markedly suppressed CRC cell proliferation and migration while inducing apoptosis. RNA sequencing analysis revealed that MTC-05 inhibited the activation of several key kinase signaling pathways, including Hippo, Wnt, PI3K/Akt, and MAPK pathways, as well as downregulated the expression of the critical target gene ZNF124. Furthermore, quantification of α-ketoglutarate (α-KG), a metabolite associated with mitochondrial function, showed that MTC-05 significantly elevated α-KG levels. Acting as a second messenger, α-KG inhibited the activation of the ZNF124/YES1/YAP1 signaling axis, thereby impeding CRC progression. In vivo experiments confirmed that MTC-05 effectively suppressed tumor growth and metastasis in CRC models without inducing cytotoxic effects. Conclusion MTC-05 exerts antitumor activity against colorectal cancer by targeting mitochondria and modulating α-ketoglutarate metabolism, which in turn suppresses the activation of the ZNF124/YES1/YAP1 signaling pathway, thereby inhibiting malignant progression.