Several investigations have also documented comparable results, particularly studies focused on the mechanisms by which the MO leaf extract (MOLE) induces tumour cell apoptosis. Multiple investigations have documented that the MOLE induces a higher production of reactive oxygen species (ROS), which in turn causes DNA fragmentation, leading to apoptosis. El-Hussieny et al., observed comparable results of increased caspase 3 production and apoptotic structural alterations, alongside a drop in average nuclear area factor readings. MO demonstrated a detrimental impact on the growth of Hep-2 cells by inducing programmed cell death by the activation of caspase 3, a hallmark of apoptosis [20] Das et al. [19] reported good dose-dependent effectiveness in their MTT assay. CAL27 and SCC15 cell lines demonstrated a substantial decline in cell survival after 24 h, with IC50 values of 17.78 µg/mL and 24.28 µg/mL for each of the groups treated with MO, respectively. Wang et al. [22] found that aqueous extracts had the most antioxidant activity in leaves, while 70% ethanol extracts exhibited the best effect in the stems, roots, and seeds of MO. In addition, MO extracts had shown strong anti-cancer activity against head and neck cancerous cells.

Abd-Rabou et al. [29, 30] observed that the nanoformulation of MO seed oil extract induced more cytotoxicity in colorectal cancer cells through mitochondrial breakdown in comparison to the free form of MO. Conversely, MO nanoformulations and MO effectively trigger mitochondrial apoptosis in HCT 116 cells, while causing insignificant harm to normal BHK-21 cells. TNF-α and HSP27 levels varied considerably across control and treatment groups, with the 125 μg/mL treatment group exhibiting the highest levels. IL-10 expression differed considerably across control and treatment groups, with the most elevated levels found in the group receiving a concentration of 500 μg/mL [24]

Immunohistochemical examination was utilized to measure IL-8 expression, and the data analysis showed that the 40 mg/kg body weight had the least level of IL-8 production in the endothelial cells of the animal model that was cancer-induced by benzo(a)pyrene [25] The MOLE can control the production of Caspase3, VEGF, and HSF1 in rodents with OSCC. Caspase-3 is a caspase executor that can be induced directly from caspases 8 and 9. The activated caspase ultimately leads to DNA fragmentation, resulting in apoptosis in cancerous cells. The Rattus norvegicus groups receiving MOLE at 3.125% and 6.25% were able to boost caspase-3 production in the cancer category. Still, the effect was enhanced in the group administered with MOLE at 9.375% [26].

Immunohistochemistry analysis demonstrated that heat shock factor-1 activity in the treatment groups was significantly lower than in the positive control group. Nevertheless, no significant variations were observed between the therapy groups [27]. Hartono et al. [28] reported a similar conclusion, with a substantial reduction in VEGF production in all treatment groups in comparison with the untreated controls. On the contrary, no significant change was found between the intervention groups.

Luetragoon et al. [21], documented that MO extracts of leaves and fractions substantially triggered apoptosis in the SCC15 cells. The findings were validated by triggering the apoptosis signalling mechanism, which resulted in dramatically increased pro-apoptotic and cleaved caspase-3 production while reducing anti-apoptotic protein Bcl-2 levels as contrasted to the control group. He found that fractions of MO leaves were capable of effectively suppressing the development of colonies and migration of cells in the SCC15 cells. Al-Asmari et al. documented that the administration of MOLE led to the interruption of the cell cycle at the stage of G2/M in tumour cells [30]. Tumour growth is characterized by the migration of cells and colony formation. The colony formation analysis was employed for examining the cell viability and growth of tumour cells determined by the propensity of individual cells to develop into cell clusters. The wound closure assays enable to make observations of cell movement in cultured cells [31]. Prior comprehensive investigations by Sreelatha et al. [7], and Sreelatha and Padma [14] demonstrated the cytotoxic impact of MOLE on KB cells along with their ability to trigger apoptosis and antiproliferative properties in KB cells. The MTT assay demonstrated that the hot water MOLE at 100 µg/mL exhibited a decrease in cell viability of around 38%. Furthermore, increasing its concentration to 200 µg/mL led to a decline in cell viability of 60%. Therefore, the MOLE effectively suppressed the KB cell growth to a degree that was contingent on the dosage.

Several morphological alterations indicative of cell death were detected in the treated cells, including shrinkage of the cytoplasmic membrane, loss of interaction with adjacent cells, the development of membrane blebs, and the occurrence of apoptotic bodies. Propidium iodide (PI), a fluorescent intercalating compound, binds to DNA and visualizes cell nuclei. In addition, PI is employed as an indicator of the degree of apoptosis in cells, as it is too large to penetrate the intact cell membrane of cells that are alive but is progressively capable of entering cells that are enduring apoptosis. Specifically, cells treated with a concentration of 200 µg/mL of MOLE exhibited nuclear shrinkage, DNA condensation, and disintegration. The study presents the potent anticancer effects of MOLE on KB cells and is accomplished by using elevating expressions of ROS within the cell and causing fragmentation of DNA [7] Additionally, several investigations have shown comparable findings [16, 32].

Berkovich et al. [33] demonstrated that the MOLE not only exhibits cytotoxic effects on Panc-1 cells but also has the potential to significantly enhance the efficacy of cisplatin when employed as a combination. The findings of Jung et al. indicate that the MOLE induces apoptosis in A549 cells by disrupting the process of gene translation [34]. The individual chemicals thiocarbamate and isothiocyanate found in MO can prevent the growth of malignant cells [35, 36] The fraction of dichloromethane has been identified to have cytotoxic effects on MCF7 cancer cells [37] Niazimincin serves as a potent chemopreventive substance in the process of carcinogenesis [38]. The extracts obtained from fruits and leaves, which include alcohol and methane, have demonstrated a noteworthy inhibition of tumour growth in animal models of melanoma [39]. Cold distilled water from Moringa, when soluble, hindered the growth of tumour cells and decreased the presence of ROS in cancerous cells [40] According to a recent computer modelling investigation, MO has been reported to contain rutin, which has the strongest binding capacity with BRAC-1, a gene associated with breast cancer [41].

The majority of the phytocomponents have demonstrated efficacy against many diseases, including cancer, which is believed to be attributed to the existence of bioactive molecules. Nano-particles developed as a result of targeted treatments, have proven to be highly efficient in a range of scientific fields owing to their minuscule dimensions. Green synthesis holds great promise for producing gold nanoparticles using indigenous vegetation and trees, like MO. The utilization of indigenous plants and trees, such as MO, in the manufacture of gold nanoparticles, has significant potential for green synthesis. The resulting gold phytonanoparticles have practical applications in cancer treatments, leading to enhanced chances of survival and improved quality of life [42] The binding energy, a crucial parameter between the protein and ligand is produced by the molecular docking process. This presents information regarding the affinity and potency of protein and ligand-receptor docking. As the binding energy decreases, the binding capacity and docking increase. The protein data that have previously been uploaded to various databases may not be identical to one another due to variations in the procedures used. Future perspectives for this arena of research involve comparing the protein structures that are acquired from various databases with various experimental models, instruments, and software. Additionally, it is imperative to evaluate the protein and gene regulation in both laboratory experiments and living organisms to confirm the computational predictions [4].

After much investigation, it has been determined that Moringa oleifera has many advantages for the human population. It is a good pharmacological choice for anti-cancer and anti-apoptotic applications because of its strong antioxidant content and diverse nutritional and phytoconstituent makeup.

Nevertheless, the present review was subjected to certain limitations. The different studies employed diverse extraction methods and solvents, potentially contributing to the disparities in findings caused by varying quantities of active chemicals in the extracted substance. Further research is required to determine any variations in the anticancer effectiveness resulting from the various extraction methods and solvents utilized. The current review included studies from in-vitro and animal models since sufficient human trials were not conducted. Future studies on human cells are required to prove the extensive potential of Moringa oleifera.