## 3 Results and Discussion
### Cell Viability  
To assess cell viability after treatment with _C. sativa_ extract, the MTS test was performed. Seven human cell lines were selected for evaluation, such as Caco-2 and DLD-1 (colon adenocarcinoma), MCF-7 (breast adenocarcinoma), SK-Mel-28 (melanoma), U87 MG (glioblastoma), U251 MG cell lines (astrocytoma) and CCD 841 CoN (colon epithelial cells). The obtained results are presented in Figure 1A.  
Among the tested cancer lines, the highest cytotoxic activity was demonstrated against Caco-2 cells (adenocarcinoma colon). The obtained IC\({}_{50}\) value was 153.54 mg/mL. Cytotoxic activity against other tumor lines was in the following order: DLD-1 (166.31 mg/mL) > MCF-7 (184.86 mg/mL) > U87 MG (210.67 mg/mL) > U251 MG (268.49 mg/mL) > SK-Mel-2 (625.94 mg/mL). At the same time, the effect of the extract on the viability of human colonic epithelial cells (CCD 841 CoN) was assessed. The obtained IC\({}_{50}\) value was 309.88 mg/mL. For the _C. sativa_ leaf methanolic extract, this is a result above the bioactive effect against the five analyzed cancer lines. Only for the SK-Mel-28 cell line was higher cytotoxic activity observed.  
\begin{table}
\begin{tabular}{c c c c} \hline \hline  & \multicolumn{3}{c}{**Anti-Diabetic and Anti-Obesity Activities**} \\ \hline  & \(\alpha\)-amylase & \(\alpha\)-glucosidase & lipase \\ \hline  & \multicolumn{3}{c}{IC\({}_{50}\) (mg/mL)} \\ \cline{2-4} _C. sativa_ leaf methanolic extract & \(1.62\pm 0.24\) & \(2.20\pm 0.22\) & \(0.72\pm 0.04\) \\ \hline \end{tabular} Values are expressed as mean \(\pm\) SD.  
\end{table}
Table 4: Anti-diabetic and -obesity activities of _C. sativa_ leaf methanolic extract.  
Figure 1: Test results for anticancer analysis. (**A**) Viability IC\({}_{50}\) values of seven cell lines after 48 h of treatment with _C. sativa_ leaf methanolic extract. (**B**) Graph showing the decrease in colony formation of colorectal adenocarcinoma (Caco-2) cells after 12 days of treatment with _C. sativa_ leaf methanolic extract. Untreated cells were selected as a control. (**C**) Representative photos of stained Caco-2 cell colonies. (**D**) Graph showing the percentage of wound closure after 12, 24 and 48 h of treatment with _C. sativa_ leaf methanolic extracts. A value of 100% means complete closure of the wound. (**E**) Representative images of cell migration after treatment with three noncytotoxic concentrations (values below IC\({}_{50}\)). Cells were analyzed at four time points (0, 12, 24, 48 h). Images were taken under an inverted microscope at 20\(\times\) magnification. All experiments were performed at least in triplicate \(n\geq 3\). Data are expressed as mean \(\pm\) SD. Significant differences (**\(p<\)0.01, ***\(p<\)0.001) were assessed using Student’s \(t\)-test and [PERSON]’s test (a–e).  
cytotoxic effect of _C. sativa_ shell extract against the same cancer line [44; 45]. On the other hand, the strongest anticancer effect of _C. sativa_ shell was demonstrated against HepG2 cells (hepatocellular carcinoma cells) (137.0 \(\upmu\)g/mL), which, according to the authors, was related to the induction of depolarization of the cell mitochondrial membrane and the modification of cell cycle phases [43; 45].