Cytotoxic Effects and Antioxidant Mechanisms of Shilajit-Loaded Chitosan Nanoparticles on HCT116 Colorectal Cancer Cells

Document Type : Original Article

Authors

1 Department of Nutrition and Food Science, Faculty of Home Economic, Al-Azhar University,

2 Professor of Genetics

Abstract

Colorectal cancer is the second leading cause of cancer deaths globally and is the most common cancer linked to obesity. This study examines Shilajit-Loaded Chitosan Nanoparticles on HCT116 Cancer Cells, organized into five groups: The first group HCT116 cells. The second group: chitosan nanoparticles-treated HCT116 cells. The third group was Shilajit extract-treated HCT116 cells. The fourth group was Shilajit-loaded chitosan nanoparticles-treated HCT116 cells. The MTT assay results demonstrated the dose-dependent cytotoxic effects of chitosan nanoparticles and shilajit-loaded chitosan nanoparticles on HCT116 cells, with IC50 values of 114.8 μg/mL and 247.5 μg/mL, respectively, compared with untreated controls. Notably, the shilajit-loaded nanoparticles exhibited minimal cytotoxicity on normal skin fibroblast cells, showing no effect up to 25 μg/mL and only a 1.64% reduction in viability at 50 μg/mL. In contrast, chitosan nanoparticles caused minimal inhibition of cell viability at various concentrations. In addition, shilajit-loaded chitosan nanoparticles dramatically decreased intracellular reactive oxygen species (ROS) and increased antioxidant enzyme compared with controls. Gene expression analysis revealed that all treatments upregulated the pro-apoptotic gene (Bax) and downregulated the anti-apoptotic gene (Bcl2) compared with controls. The extract-loaded nanochitosan exhibited the highest Bax expression, whereas shilajit resulted in lower Bcl2 expression. Interestingly, the treatments increased the expression of the oncogenes c-MYC and KRAS in colon cancer cells, with the highest expression observed in the shilajit-loaded nanochitosan group. The combination of shilajit and chitosan nanoparticles presents a promising and biocompatible delivery system for targeted cancer therapy, with minimal harm to healthy cells and potential synergy in treatment strategies.

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Articles in Press, Corrected Proof
Available Online from 08 July 2025
  • Receive Date: 25 March 2025
  • Revise Date: 26 April 2025
  • Accept Date: 02 July 2025