Bioactivity of Mineral-Enriched Organic Carrots: Anticancer Effects in HT-29 Cells and Anti-Inflammatory Activity in Mice Splenocytes

Carrot (Daucus carota) is a globally important root vegetable with diverse biological activities. Currently, organic cultivation places a greater focus on ecosystem health by using natural organic fertilisers and organic pesticides, such as animal and plant-based fertilisers and natural insecticides, reducing reliance on chemical substances, promoting soil health and ecological balance, and providing healthier and environmentally friendly agricultural products. However, research on the anti-inflammatory properties of carrots, especially comparing traditionally cultivated and organically grown varieties, remains limited. Therefore, this study examined the anticancer and anti-inflammatory properties of carrots cultivated using Natural Dream Cultivation (NC), an organic farming method enhanced with mineral supplementation. The inhibitory effect of NC carrot extracts on HT-29 human colon cancer cells was assessed via MTT assay, while qPCR was used to evaluate mRNA expression of cell cycle- and apoptosis-related genes. Nitric oxide (NO) concentration was measured using the Griess method. In LPS-induced mouse splenocytes, levels of inflammatory cytokines were determined by enzyme-linked immunosorbent assay, and natural killer (NK) cell activity was evaluated via LDH release assay. Results showed that NC significantly suppressed cancer cell proliferation. It up-regulated the expression of cell cycle arrest genes (p53, p21) and pro-apoptotic genes (Bim, Bad, Bax, Bak, caspase-9, caspase-3), while down-regulating anti-apoptotic genes (Bcl-2, Bcl-xL). Furthermore, NC treatment inhibited NO production and reduced the release of inflammatory cytokines (TNF-α, IL-6, IL-1β, IFN-γ, IL-12) in LPS-stimulated splenocytes. NC also promoted NK cell activation. By elucidating the inhibitory effects on cancer cells and modulation of inflammatory responses, this study reveals potential mechanisms behind the anticancer and anti-inflammatory activities of carrots. The innovative mineral-enhanced organic cultivation method presented here offers new possibilities for developing carrots as a functional food candidate supported by in vitro and ex vivo bioactivity relevant to cancer- and inflammation-associated pathways. These findings may inform the future design of dietary research hypotheses and preclinical studies, pending validation in animal models and clinical settings.