Assessing the Therapeutic Impacts of HAMLET and FOLFOX on BRAF-Mutated Colorectal Cancer: A Study of Cancer Cell Survival and Mitochondrial Dynamics In Vitro and Ex Vivo.

Background and Objectives: Colorectal cancer (CRC) is a major global health challenge. The BRAF V600E mutation, found in 8-12% of CRC patients, exacerbates this by conferring poor prognosis and resistance to therapy. Our study focuses on the efficacy of the HAMLET complex, a molecular substance derived from human breast milk, on CRC cell lines and ex vivo biopsies harboring this mutation, given its previously observed selective toxicity to cancer cells. Materials and Methods: we explored the effects of combining HAMLET with the FOLFOX chemotherapy regimen on CRC cell lines and ex vivo models. Key assessments included cell viability, apoptosis/necrosis induction, and mitochondrial function, aiming to understand the mutation-specific resistance or other cellular response mechanisms. Results: HAMLET and FOLFOX alone decreased viability in CRC explants, irrespective of the BRAF mutation status. Notably, their combination yielded a marked decrease in viability, particularly in the BRAF wild-type samples, suggesting a synergistic effect. While HAMLET showed a modest inhibitory effect on mitochondrial respiration across both mutant and wild-type samples, the response varied depending on the mutation status. Significant differences emerged in the responses of the HT-29 and WiDr cell lines to HAMLET, with WiDr cells showing greater resistance, pointing to factors beyond genetic mutations influencing drug responses. A slight synergy between HAMLET and FOLFOX was observed in WiDr cells, independent of the BRAF mutation. The bioenergetic analysis highlighted differences in mitochondrial respiration between HT-29 and WiDr cells, suggesting that bioenergetic profiles could be key in determining cellular responses to HAMLET. Conclusions: We highlight the potential of HAMLET and FOLFOX as a combined therapeutic approach in BRAF wild-type CRC, significantly reducing cancer cell viability. The varied responses in CRC cell lines, especially regarding bioenergetic and mitochondrial factors, emphasize the need for a comprehensive approach considering both genetic and metabolic aspects in CRC treatment strategies.

Species-Specific Plant-Derived Nanoparticle Characteristics.

Medicinal and agricultural plants contain numerous phytochemical compounds with pronounced biological effects on human health. They are known to encapsulate most of their characteristic bioactive compounds within membranous elements of intercellular communication known as exosomes. These nanovesicles serve as capsules protecting their biological activity and improving their penetration into the tissue. Therefore, the application of plant exosome preparations holds considerable potential for cosmetics and pharmacy, but the quality and consistency of plant material for exosome isolation is of critical importance. Therefore, in this study, we aimed to evaluate yield, size distribution patterns, and antioxidant properties between nanovesicle preparations of the following portfolio of medicinal plants: Kalanchoe daigremontiana, Artemisia absinthium, Hypericum perforatum, Silybum marianum, Chelidonium majus, and Scutellaria baicalensis. Results showed that nanoparticle yield, size distribution, and antioxidant activities were specific to plant species. Compared to other plants, nanoparticle preparations from Artemisia absinthium were distinguished by remarkably higher yield and concentration, while the highest antioxidant activity of plant-derived nanoparticle preparations per weight and per particle was determined to occur in Chelidonium majus and Hypericum perforatum samples. Results showed no significant correlation in DPPH (2-diphenyl-1-picrylhydrazyl) free radical scavenging activity and FRAP (ferric reducing antioxidant power) between plant material and nanoparticle preparations. More detailed biochemical analysis of exosome preparations is necessary to validate their biological activity and its relation to source plant cells.