## 2 Results and Discussion
### Anticancer Potential  
Various concentrations of MH were tested against three cancer cell lines (HCT-116, MCF-7, A549), and their cytotoxic effects were observed using MTT assays. The percentage of cell viability at different concentrations of the MH sample and the IC\({}_{50}\) values were determined (Table 5).  
Anticancer activity of MH against tested cell lines are surmised in Figure 2. A dose-dependent anti-cancer potential was observed, as well as a substantial anticancer potential against the MCF-7 and A549 cell lines, and IC\({}_{50}\) values were as 9.05 and 9.37 mg/mL, respectively. Aside from its antibacterial and antioxidant characteristics, honey has been found in recent research to have features that could slow the multiplication of malignant cells [45; 53; 54]. The anti-cancerous activity can occur via the adoption of one of the following routes: (1) depolarizing the mitochondrial membrane of cancer cells, hence causing apoptosis, (2) cyclooxygenase-2 inhibition by components, such as flavonoids, (3) releasing cytotoxic H\({}_{2}\)O\({}_{2}\) and (4) scavenging the reactive oxygen species (ROS). It has also been associated with the phytochemical compounds [55]. On human breast cancer (MCF-7), colorectal carcinoma (CT26) and murine melanoma (B16.F1), MH has been shown to have a dose and time-dependent anti-proliferative effect [37]. It is primarily  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Tested Bacteria** & **Mean** & **\% Inhibition** \\ \hline _B. subtilis_ & Control & 0.76 \(\pm\) 0.01 & 67.13 \\
1/2 MIC & 0.25 \(\pm\) 0.01 & 67.13 \\ \hline _S. aureus_ & Control & 0.84 \(\pm\) 0.01 & 56.52 \\
1/2 MIC & 0.36 \(\pm\) 0.05 & 56.52 \\ \hline _E. coli_ & Control & 0.72 \(\pm\) 0.03 & 48.28 \\
1/2 MIC & 0.37 \(\pm\) 0.01 & 48.28 \\ \hline _P. aeruginosa_ & Control & 0.87 \(\pm\) 0.01 & 42.78 \\
1/2 MIC & 0.50 \(\pm\) 0.04 & 42.78 \\ \hline \hline \end{tabular} All results are presented as the mean \(\pm\) standard deviation.  
\end{table}
Table 4: Antibiofilm activity of the manuka honey against biofilm-forming bacterial species.  
\begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline
**Tested Cell** & **Percentage of Inhibition** & \multicolumn{6}{c}{**Honey Concentrations (\%)**} & \multirow{2}{*}{**IC\({}_{50}\)**} \\ \cline{3-4}
**Lines** & **and Cell Viability** & **2** & **4** & **6** & **8** & **10** \\ \hline \multirow{2}{*}{HCT-116} & Inhibition rate (\%) & 18.52 & 31.78 & 42.03 & 54.34 & 64.73 & 48.65 \\ \cline{2-4}  & Cell viability (\%) & 81.47 & 68.21 & 57.96 & 45.65 & 35.26 & \\ \hline \multirow{2}{*}{MCF-7} & Inhibition rate (\%) & 12.53 & 23.22 & 34.04 & 42.23 & 57.81 & \multirow{2}{*}{9.05} \\  & Cell viability (\%) & 87.46 & 76.77 & 65.95 & 57.76 & 42.18 & \\ \hline \multirow{2}{*}{A549} & Inhibition rate (\%) & 9.73 & 15.86 & 28.31 & 37.43 & 49.89 & \multirow{2}{*}{9.37} \\  & Cell viability (\%) & 90.26 & 84.13 & 71.68 & 62.56 & 50.10 & \\ \hline \hline \end{tabular} \(\text{IC}_{50}\) results are presented as the mean of three reads. HCT-116: colon cancer cells; MCF-7: breast cancer cells; A549: lung carcinoma epithelial cells.  
\end{table}
Table 5: Percentage of inhibition, cell viability, and IC\({}_{50}\) values of tested manuka honey against various cancer cell lines.  
mitochondrial membrane apoptosis [56]. The initiator caspase-9 is triggered, which causes the executioner caspase-3 to be activated [55]. It also promotes apoptosis by activating poly(ADP-ribose)polymerase protein (PARP), which causes the loss of Bcl-2 expression and fragmentation of DNA [57]. In a mouse melanoma model, MH significantly lowered overall tumor volume and boosted death of these cells [58]. A study by [PERSON] and colloques used rats suffering from inflammatory bowel disease, and after applying HM, colonic inflammation was decreased, lipid peroxidation level was lowered, and the antioxidant activities were increased [55]. These findings justify the observed cytotoxicity potential of the tested MH sample.