Title:Therapeutic Effects of Crocin Nanoparticles Alone or in Combination with Doxorubicin against Hepatocellular Carcinoma In vitro
Volume: 25
Issue: 3
Author(s): Noha S. Basuony, Tarek M. Mohamed*, Doha M. Beltagy, Ahmed A. Massoud and Mona M. Elwan
Affiliation:
- Department of Biochemistry, Faculty of Science, Tanta University, Tanta, Egypt
Keywords:
Hepatocellular carcinoma, crocin, crocin nanoparticles, doxorubicin, apoptosis, autophagy.
Abstract:
Objective: Crocin (CRO), the primary antioxidant in saffron, is known for its anticancer properties.
However, its effectiveness in topical therapy is limited due to low bioavailability, poor absorption, and low physicochemical
stability. This study aimed to prepare crocin nanoparticles (CRO-NPs) to enhance their pharmaceutical
efficacy and evaluate the synergistic effects of Cro-NPs with doxorubicin (DOX) chemotherapy on two cell
lines: human hepatocellular carcinoma cells (HepG2) and non-cancerous cells (WI38).
Methods: CRO-NPs were prepared using the emulsion diffusion technique and characterized by transmission
electron microscopy (TEM), scanning electron microscopy (SEM), Zeta potential, and Fourier transform infrared
spectroscopy (FT-IR). Cell proliferation inhibition was assessed using the MTT assay for DOX, CRO, CRO-NPs,
and DOX+CRO-NPs. Apoptosis and cell cycle were evaluated by flow cytometry, and changes in the expression
of apoptotic gene (P53) and autophagic genes (ATG5 & LC3) were analyzed using real-time polymerase chain
reaction.
Results: TEM and SEM revealed that CRO-NPs exhibited a relatively spherical shape with an average size of 9.3
nm, and zeta potential analysis indicated better stability of CRO-NPs compared to native CRO. Significantly
higher antitumor effects of CRO-NPs were observed against HepG2 cells (IC50= 1.1 mg/ml and 0.57 mg/ml)
compared to native CRO (IC50 = 6.1 mg/ml and 3.2 mg/ml) after 24 and 48 hours, respectively. Annexin-V assay
on HepG2 cells indicated increased apoptotic rates across all treatments, with the highest percentage observed in
CRO-NPs, accompanied by cell cycle arrest at the G2/M phase. Furthermore, gene expression analysis showed
upregulation of P53, ATG5, and LC3 genes in DOX/CRO-NPs co-treatment compared to individual treatments.
In contrast, WI38 cells exhibited greater sensitivity to DOX toxicity but showed no adverse response to CRONPs.
Conclusion: Although more in vivo studies in animal models are required to corroborate these results, our findings
suggest that CRO-NPs can be a potential new anticancer agent for hepatocellular carcinoma. Moreover, they
have a synergistic effect with DOX against HepG2 cells and mitigate the toxicity of DOX on normal WI38 cells.
