# Increased Anticancer Activity of Organic Kimchi with Starters Demonstrated in HT-29 Cancer Cells
## 4 Discussion
This is summarized as cell death due to cell morphology and internal biochemical changes, which include cell rupture, cell membrane changes, nuclear fragmentation, chromatin condensation, and chromosome cleavage, and ends with the cells being phagocytosed and processed by other cells [50]. The first link between apoptosis and cancer began in the late 1980s, when it was understood that the Bcl-2 gene specifically blocks B-cell apoptosis in lymphomas [51]. The mitochondrial pathway of apoptosis is the major mechanism of physiological cell death, which is initiated by the pro-apoptotic members of the Bcl-2 family that cause mitochondrial outer-membrane permeabilization (MOMP). Pro-apoptosis, which induces the release of cytochrome c from mitochondria, and anti-apoptosis, which inhibits the release, play a role in apoptosis by interacting with each other [52]. Anti-apoptosis genes include Bcl-2 and Bcl-xL, which inhibit apoptosis and pro-apoptosis genes, such as Bax and Bak, which are downstream effectors that directly induce and promote apoptosis [53]. In addition, the pro-apoptotic BH3-only protein is an upstream member of the chain reaction, and when activated, suppresses anti-apoptosis and activates pro-apoptosis factors, such as Bax and Bak, to induce mitochondrial apoptosis [54]. [PERSON] et al. [4] reported that kimchi supplemented with functional additives down-regulated Bcl-2 and up-regulated pro-apoptotic protein expression in HT-29 cells. Furthermore, [PERSON] et al. [9] confirmed that kimchi with starters increased the mRNA expression of Bax and decreased the expression of Bcl-2, making it more effective in regulating apoptosis-related factors. In this study, the mRNA expression of anti-apoptotic Bcl-2 was decreased in HT-29 cells by treatment with OAK, an organic functional kimchi, and the mRNA expressions of pro-apoptotic Bak and BH3-only proteins Bim and Bad were up-regulated (Figure 5). In addition, the measurement of protein expression levels (Figure 6) confirmed that the expression of Bcl-2 decreased and the expression of Bax increased. The caspase-cascade system plays an important role in the induction, transduction, and amplification of intracellular apoptotic signals. Caspases are classified according to their roles into initiator (caspases 2, 8, 9, and 10), executioner (caspases 3, 6, and 7), and inflammatory caspases (caspases 1, 4, and 5) [55]. Caspases 3, 7, and 9 have been reported to play a central role in the apoptotic process. When caspase 9 is activated, it cleaves and activates caspases 3/7 to further cleave other endogenous substrates and induce apoptosis [56]. Caspase 3 degrades intracellular structural and functional proteins and causes apoptosis [57]. In this study, the expression levels of caspases 3 and 9 were the highest in OAK, and OAK regulated the expression levels of caspase family members, resulting in high apoptotic activity in HT-29 cells. These results are similar to those of [PERSON] et al. [37], in which the significant up-regulation of caspases 3 and 9 was observed in aged kimchi. These results show that kimchi supplemented with a functional recipe of nF1, starters, and organic ingredients can play an important anticancer role by regulating the expression of cell cycle arrest, apoptosis, and caspase-related genes. Considering the difference between kimchi samples prepared with different ingredients and lactic acid bacteria, it seems that the functional recipe of OAK prepared with the abovementioned ingredients improves the fermentation quality and antioxidant and anticancer effects of kimchi. The kimchi recipes prepared in this way were the best anticancer kimchi recipes confirmed after 20 years of research in this laboratory. Therefore, OAK seems to be an ideal kimchi for the development of premium organic anticancer kimchi.