Quercetin and 5-Fu Loaded Chitosan Nanoparticles Trigger Cell-Cycle Arrest and Induce Apoptosis in HCT116 Cells via Modulation of the p53/p21 Axis

Abstract

Nanoparticles (NPs) are encapsulating agents that exist in the nanometer range. They can be classified into different classes based on their properties, shapes, or sizes. Metal NPs, fullerenes, polymeric NPs, ceramic NPs, and luminescent nanoporous hybrid materials are only a few examples. This study explored the anticancer potential of quercetin and 5-fluorouracil-encapsulated chitosan nanoparticles (CS-5-FU-QCT NPs). The nanoparticles were prepared by ionic gelation, and their efficacy and mechanism of action were examined. CS-5-FU-QCT NPs were characterized using dynamic light scattering (DLS), atomic force microscopy (AFM), UV−visible spectroscopy, and Fourier transform infrared spectroscopy (FTIR); cytotoxicity was analyzed using an MTT assay. Cells were treated with CS-5-FU-QCT NPs and incubated for 12, 24, and 36 h, and apoptosis analysis (using Annexin V/FITC), cellcycle analysis, Western blotting, and confocal microscopic analysis were performed. Biophysical analysis revealed that the CS-5-FU-QCT NPs fall in the range of 300−400 nm with a near-spherical shape. The in vitro drug release profile indicates sustained release of drugs over a period of about 36 h. The cytotoxicity of CS-5-FU-QCT NPs was more prominent in HCT116 cells than in other cancer cells. This particular nanoformulation caused G0/G1 phase cell-cycle arrest in HCT116 cells and induced intracellular ROS generation, thereby causing apoptosis. It also downregulated Bcl2, cyclin D1, and Cdk4 and upregulated BAX, p53, and p21, causing cell-cycle arrest and apoptosis. In summary, CS-5-FU-QCT NPs hindered proliferation of HCT116 cells via ROS generation and altered the expression of key proteins in the p53/p21 axis and apoptotic machinery in a time-dependent manner.

Quercetin: A Potential Polydynamic Drug

“…In 2023, Das Saha et al [141] examined the anticancer potential of quercetin and 5fluorouracil-encapsulated (5-Fu) chitosan nanoparticles, since chitosan has been intensively investigated and used as a carrier in polymeric nanoparticles for drug delivery in both in vitro and in vivo models [142]. 5-Fu is already a chemotherapeutic drug approved by the FDA for treating various types of cancers [143], and quercetin has the ability to express its anticarcinogenic properties via the modification of intracellular signal transduction and the inhibition of cancer-activating enzymes [144].…”

“…The products of this reaction have antioxidant activity and may be used as novel drug leads [140]. In 2023, Das Saha et al [141] examined the anticancer potential of quercetin and 5fluorouracil-encapsulated (5-Fu) chitosan nanoparticles, since chitosan has been intensively investigated and used as a carrier in polymeric nanoparticles for drug delivery in both in vitro and in vivo models [142]. 5-Fu is already a chemotherapeutic drug approved by the FDA for treating various types of cancers [143], and quercetin has the ability to express its anticarcinogenic properties via the modification of intracellular signal trans-Scheme 9.…”

“…The results of the performed MTT assays indicated a higher cytotoxic potential of CS-5Fu-QCT nanoparticles in HCT116 cancer cells and no toxicity on the tested normal kidney cells compared to quercetin alone, meaning that this type of NP may be a very effective anticancer agent against colorectal carcinoma, with minimum to no side effects. The CS-5Fu-QCT NPs' possible mechanism of action works by causing G0/G1 phase cell-cycle arrest in HCT116 cells and altering the expression of pivotal proteins in the p53/p21 pathway, thus initiating cell apoptosis [141].…”