Cytotoxic effect of magnetic iron oxide nanoparticles conjugated with thymol on liver cancer cell line and evaluation of caspase-8 gene expression

Subject Areas : cellular and molecular bilology

Ali Salehzadeh ^{1 *} , Yasaman Alizadeh Kolangestani ²

1 - Associate professor, Department of Biology, Ra.C., Islamic Azad University, Rasht, Iran
2 - MSc, Department of Biology, Tabriz Branch, Islamic Azad University, Tabriz, Iran

Received: 2025-06-24 Accepted : 2025-07-29 Published : 2025-06-01

Keywords: Iron oxide, Thymol, Liver cancer, Flow cytometry,

Abstract :

Introduction: The application of magnetic nanoparticles for effective drug delivery to cancer tissues has gained attention. This study was conducted to determine the anticancer effects of magnetic iron oxide nanoparticles functionalized with glucose and conjugated with thymol (Fe₃O₄@Glu-Thymol NPs) on a liver cancer cell line.

Materials and Methods: Physicochemical assays including FT-IR, XRD, EDS, scanning electron microscopy, DLS, and zeta potential were performed to determine the functional groups, crystal structure, shape, size, and surface charge of the nanoparticles. The cytotoxic effects of nanoparticles on the liver cancer cell line (HepG2) and normal cell line (HDF) were performed using MTT assay. Flow cytometry was used to determine the percentage of apoptotic cells and real-time PCR was used to assess the expression of the caspase-8 gene.

Results: Fe₃O₄@Glu-Thymol NPs had a spherical shape, diameter of less than 60 nm, a surface charge of -13.5mV, and a hydrodynamic diameter of 515nm. The nanoparticles exhibited concentration-dependent toxicity for liver cancer cells, and their IC₅₀ in cancer and normal cell lines was 67.5 and 175 μg/mL, respectively. Flow cytometry showed that Fe₃O₄@Glu-Thymol NPs caused a significant increase in the percentage of early and late apoptosis, and the expression of the caspase-8 gene in treated cells increased by 2.45-fold (p<0.001).

Conclusion: The results of this study indicate the effective inhibitory effects of Fe₃O₄@Glu-Thymol NPs on liver cancer cells, which can be helpful in the development of effective magnetic nanodrugs for targeted drug delivery to liver cancer tissues.

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