## 3 Results and Discussion
### Analysis of Apoptosis Using the Muse(r) Flow Cytometer
A higher expression of these proteins was observed after treatment with the juice of young shoots subjected to in vitro gastrointestinal digestion and absorption in comparison to that after treatment with the juice of red cabbage subjected to the same processes (Figures 3 and 4; Supplementary Figures S3 and S4). Although a great number of research dealing with the anticarcinogenic activity of single components have been conducted, only a few studies were performed using whole Brassica vegetables, including red cabbage and in particular their young shoots. Research by [PERSON] et al. [29] on red cabbage extract may support our findings regarding the activation of different apoptotic pathways in cancer cells. They reported that red cabbage extracts up-regulated the expression of caspase-7, -8, -9, and the \(Bax\) gene, and also downregulated the expression of the Bcl-2 gene in both HeLa and HepG2 cells. Red cabbage extract was also found to induce apoptosis in the human cancer cells via caspase-dependent pathway [42]. [PERSON] and [PERSON] [43] investigated that the red cabbage extract induced cell death in MDA-MB-231 human breast cancer cells, decreased the expression of Bcl-2 gene, and increased the expression of \(Bax\) and caspase-3 genes. In our previous publication, we showed that the young shoots of red cabbage, from which the juice was obtained for the study described in the current manuscript, contain many glucosinolates, polyphenols, vitamin C, and carotenoids [10; 11]. Glucosinolates may break down in plant material during processing through endogenous enzyme myrosinase action, or within the gastrointestinal tract through the action of commensal microflora. A proportion of these breakdown products enter the epithelial cells of the gastrointestinal mucosa, undergo metabolism, enter to the circulation, and then undergo further metabolism in the liver [44]. Cruciferous sporus, including young shoots of red cabbage, could be a good dietary source of aliphatic glucosinolates, glucoraphanian being one of the most abundant, and its derivative-sulforaphane, which was reported to induce apoptosis in the human colon cancer cell line HT29 in the intrinsic pathway [45]. Moreover, sulforaphane up-regulated the \(Bax\) protein in LNCaP cells, down-regulated the expression of Bcl-2 in DU145 cells, and inhibited NF-\(\kappa\)8 transcriptional activity in PC-3 [16; 46; 47]. Another GLS present in the studied vegetable is glucoberin (precursor of the isothiamateiberin), which has beneficial effects on oxidative stress and cancer prevention [7; 10]. A high concentration of total indole GLS, represented by glucobrassicins, neoglucobrassicin, and 4-methoxyglucobrassicin, was observed in young shoots [10; 48]. [PERSON] and [PERSON] [49] demonstrated that indole-3-carboniol (I3C; derived from the breakdown of glucobrassicin) regulated many genes involving in cell cycle regulation, proliferation, and other cellular processes, suggesting the pleiotropic effects of this compound on the prostate cancer cell line. [PERSON] et al. [50] and [PERSON] et al. [51] reported that I3C was an effective agent in the inhibition of cell growth and induction of programmed cell death of PC-3 and of DU145 and LNCaP prostate cancer cells, respectively. The breakdown product of the 4-methoxyglucobrassicin (4-methoxyindole-3-carboniol) might play a role as a chemo-preventive agent, inhibiting cell growth and causing cell death of human cancer cells in vitro [52]. Phenolic extracts and single polyphenols from different plant foods have been studied in a number of cancer cell lines representing different developmental stages of cancer. Polyphenols exert their biological actions by different but complementary mechanisms. A study demonstrated the anti-proliferating effects of polyphenols, alone or in combination, on cancer cells lines, like breast (T47D, MCF7, MDA-MB-231) and prostate (DU145, PC3, LNCaPP), in a time and dose-response manner [53].