## 4 Evaluation of Biological Properties
### Anticancer Activity  
Several works have shown that daucosterol possesses important anticancer activity on various tumor cell lines and could be considered one of the novel pharmacological treatment strategies for cancer (Table 4). [PERSON] et al. [36] isolated this molecule from _Salvia sahendica_ to assess its apoptotic and anti-proliferative activity against human breast adenocarcinoma MCF-7 cells using MTT assay, lactate dehydrogenase (LDH) leakage assay, and flow cytometry. They found that daucosterol inhibits cell proliferation and induces cytotoxicity in MCF-7 cells, including PARP proteolytic activity, DNA fragmentation, and cell morphological changes. Indeed, at 80 \(\mu\)M, maximum inhibition of cell growth and survival was approximately 60%. Anti-cancer drug mechanisms move from the subcellular to the molecular level by increasing the levels of the pro-apoptotic proteins (Bax and Bcl2) and decreasing the Bcl-2/Bax ratio, upregulating the PTEN gene to inhibit the PI3K/Akt pathway, inducing the loss of mitochondrial membrane potential, and by Cyt c release in breast cancer cells [110, 112, 113]. On the other hand, [PERSON] et al. [110] assessed the anticancer effect of daucosterol against human breast cancer cell line MCF-7 and gastric cancer cell lines MGC803, BGC823, and AGS. The results demonstrated that this compound exerts an important antiproliferative activity against the studied cell lines, with IC\({}_{50}\) values of 16.95, 19.96, 3.13, and 24.19 \(\mu\)M, respectively. This effect was induced by autophagy in a ROS-dependent manner. [PERSON] et al. [38] recorded an IC\({}_{50}\) value of 26.6 \(\mu\)M on human colon cancer cell line HCT-116. Daucosterol can repress cell migration and cell invasion in these cells and cause cell death by cell-cycle arrest and apoptosis. Indeed, it significantly inhibited the proliferation of human lung cancer cell line A549, with an ICvalue of 17.46 \(\upmu\)g/mL targeting multiple checkpoints, including cell-cycle arrest at the G\({}_{2}\)/M phase and induction of cell apoptosis [112]. Using MCF-7, MDA-MB-231, and 4T1 breast cancer cells, [PERSON] et al. [37] revealed that daucosterol exhibits antitumor activities, with an IC\({}_{50}\) = 53.27 \(\upmu\)g/mL on MCF-7 and IC\({}_{50}\) > 1000 on MDA-MB-231 and 4T1. This molecule also inhibited tumor growth in vivo using MCF-7 xenografts in nude mice by decreasing the expression of Bcl-xl, Bcl-2, and XIAP, increasing Bax, Bad, inducing the activation of caspase-dependent apoptosis in tumor tissues, and inactivation of the upstream PI3K/Akt/NF-kB pathway. Moreover, [PERSON] et al. [114] reported that daucosterol inhibits cell proliferation, induces cell-cycle arrest, and promotes autophagy-dependent apoptosis in human prostate cancer cell lines (PC3 and LNCaP) via activation of JNK signaling. Indeed, this phytosterol strongly inhibited the growth of human lung cancer cell line A549, with an IC\({}_{50}\) value of 20.9 \(\upmu\)M, by increasing ROS levels and promoting intrinsic apoptotic cell death of A549 cells. This effect was mediated by increased expression of caspase-3, caspase-9, and Bax, PARP inactivation, Cyt c release, and diminished expression of bcl-2 protein (Figure 2) [26]. Recently, [PERSON] et al. [25] evaluated the antiproliferative effect of daucosterol extracted from sweet potatoes on three breast cancer cell lines (MCF-7, MDA-MB-231, and 4T1) and nontumorigenic breast epithelial cell line MCF-10A using MTT assay. The results showed that this compound inhibited the proliferation of breast cancer MCF-7 cells by inactivating the phosphoinositide 3-kinase/protein kinase B pathway, but had only weak effects on the proliferation of MDA-MB-231, 4T1, and MCF-10A cells. [PERSON] and coworkers [24] investigated antitumor effects in vivo using the environmental carcinogen 7,12-dimethylbenz[_a_]anthracene (DMBA). The results demonstrated that daucosterol reduced, at all doses, tumor volume as well as proteins and cancer antigen 1.