Assessment the Effect of Ficus carica Leaf Extract on B16F10 Melanoma Cancer Cells: The Roles of p53, Caspase-3 & Caspase-9 on Induction of Intrinsic Apoptosis Cascade

Abstract

Background: Melanoma is the most serious kind of skin cancer which has significantly increased in recent decades, and the importance of its primary treatment is increased widely. Ficus carica leaves have various therapeutic impact such as anti-inflammatory, anti-proliferative and apoptotic activity. Objectives: Hence, regarding the F. carica effect on the treatment of various diseases, the present research was conducted to identify the effect of methanolic extract of F. carica leaf on apoptosis induction in B₁₆F₁₀ melanoma cancer cells. Methods: Cell survival was estimated by MTT assay after treatment of B₁₆F₁₀ cells in various concentrations of F. carica leaf extract (150, 250, 350, 450, 550, 650, 750 and 850 μg /mL) for 24 and 48 h. Cell apoptosis was analysed by AO/PI and DAPI staining, Annexin V/Propidium Iodide flow cytometry. Moreover, Real-time PCR was utilized to evaluate the expression of apoptotic genes including p53, Bax, caspase-3 and caspase-9 genes. Results: MTT assay results indicated that methanolic extract of F. carica leaf prevented the proliferation of B₁₆F₁₀ cells in a dose and time dependent manner. AO/PI staining results showed an elevation in apoptotic cells in treated groups and DAPI indicated that F. carica extract resulted in chromatin condensation and fragmentation. Annexin V revealed the increasing percentage of apoptotic cells after treatment. In addition, the up-regulation of apoptotic genes confirmed the apoptosis inducing potential of F. carica leaves in B₁₆F₁₀ cells by Real-time PCR. Conclusions: Thus, methanolic extract of F. carica leaves could be suggested as an effective anti-cancer agent for further studies on melanoma cancer. Keywords Cytotoxicity Apoptosis Anti-proliferative Melanoma Cancer Ficus carica