Investigation of Proapoptotic Effect of Digera muricata Leaf Extract against the Skin Cancer Cell Line (B16-F10)

Abstract

Introduction: Skin cancers, such as melanoma, basal cell carcinoma, and squamous cell carcinoma, frequently begin as changes in the skin. Cancer research continues to focus on finding tumor-selective and new anticancer drugs with fewer adverse effects. Digera muricata is a medicinal herb in the Amaranthaceae family that has antibacterial, antifungal, free radical scavenging function, anti tumor, and other valuable medicinal properties.
 Aim: To investigate the pro-apoptotic effect of Digera muricata leaf extract against the skin cancer cell line.
 Materials and Methods: MTT assay was used to determine the viability of B16-F10 cells treated with different concentrations (20-200μg) of Digera muricata leaf extract. Phase contrast microscopy was used to examine the morphological changes. In addition, the mode of cell death was assessed using AO/EtBr dual staining and observed under a fluorescence microscope. Statistical analysis was performed, keeping the level of significance at p<0.05.
 Results: The MTT assay revealed that the ethanolic extract of Digera muricata leaf had significant cytotoxic and apoptotic potency against the B16-F10 skin cancer cell line, which was validated by significant morphological alterations under phase contrast microscope after 24 hours of treatment. AO/EtBr dual staining results clearly showed the Digera muricata leaf extract treatment induced the early apoptotic cells with bright orange areas of condensed or fragmented chromatin in the nucleus. Late apoptotic cells showing uniform bright red nucleus.
 Conclusion: Within the limits of the analysis, it can be inferred that the leaf extract of Digera muricata was cytotoxic and triggered cancer cell apoptosis at a concentration of 50 μg/ml within 24 hours. More research is needed to understand the cytotoxicity mechanisms of this plant extract.