## 4 Discussion
Besides its rich alfa- and beta-carotene contents, the root contains a wide range of bioactive compounds, including chlorogenic acids [35], quercetin, luteolin, kaempfer, myricetin [36], cyaniding, pelargonidin, and peonidin [37] as phenolic compounds; falcarinol, falcarindiol, and falcarindiol-3-acetate [38] as polyacetylenes; and vitamin C [34]. According to research, carotenoids have been clinically studied as an anticancer agent and found effective in animal models and humans. Beta-carotene is a very strong antioxidant compound because of its 11 conjugated doublebonds and a \(\beta\)-ring at each end of the chain, which may neutralize free radicals in lipophilic environments, including membranes [35]. Extracts obtained through hexane from red carrots have been found to have cytotoxic activity against human breast cancer cell lines (MCF-7) [39]. Similarly, ethanol or acetone extracts from black carrots have been shown to have cytotoxic activity against the same cancerous cell line. Therefore, the carrot samples in this study could potentially serve as a source for the extraction of carotenoids, which might be developed as targeted agents for cancer therapy in the future. The inhibition of cancer cell growth is one of the key mechanisms underlying anticancer activity. Cancer cell control often involves regulating the cell cycle and promoting apoptosis [40]. p53, a tumor suppressor protein, is essential for preserving genomic stability and initiating cell apoptosis. p53 is activated in response to DNA damage or other stress conditions and initiates a series of responses, such as cell cycle arrest, DNA repair, and apoptosis induction [41]. p21, a cell cycle regulatory protein, is a downstream target gene of p53. p21 can inhibit cell cycle progression, prevent cell proliferation and DNA replication, and guide cells to apoptosis [42]. Bcl-2 family proteins associated with apoptosis include Bim, Bad, Bcl-2, Bcl-xl, Bax, and Bak. Bim and Bad belong to the BH3-only protein group, which is important in regulating cell apoptosis. Bim and Bad inhibit anti-apoptotic members of the Bcl-2 family, such as Bcl-2 and Bcl-xL, while releasing or activating pro-apoptotic proteins, such as Bax and Bak, within the cell. These proteins operate directly on the outer mitochondrial membrane, increasing the permeability of the mitochondrial outer membrane and the release of apoptotic factors, such as cytochrome C, eventually triggering cell apoptosis [43]. Caspase-9 and caspase-3 are key cysteine aspartate-specific protease family members that act as apoptosis executioners. Caspase-9 activation can activate caspase-3, resulting in a cascade of apoptosis-related reactions that eventually lead to cell apoptosis [44]. According to the study in [45], the water extract of purple carrot anthocyanins effectively inhibited the growth of HT-29 cells [46], whereas the extract of black carrot exhibited inhibitory effects on the proliferation of various human cancer cell lines, including MDA-MB-231 (human breast cancer), HT-29, and PC-3 (human prostate cancer) [47,48]. In this study, carrots demonstrated growth inhibitory effects on HT-29 cancer cells, particularly OC and NC. Moreover, NC exhibited improved cell cycle and apoptosis-related gene regulation in HT-29 cells compared with CC and OC. These results suggest that mineral-enriched organic cultivation improves carrot control over cancer cells and that a diet rich in NC may potentially prevent cancer growth. Inflammation is the body's immune response to injury, infection, or other stimuli, and it involves the production and interaction of various inflammatory mediators. Nitric oxide (NO) is a signaling molecule that plays a vital role in the pathogenesis of inflammation, and abnormal NO production is an important marker of the inflammatory response to LPS treatment [49,50]. Tumor necrosis factor-alpha (TNF-\(\alpha\)) is a central cytokine in the inflammatory response, and neutralizing TNF has been one of the most effective treatments for chronic inflammation and autoimmune diseases [51].