Pengaruh Pemberian Ekstrak Buah Andaliman (Zanthoxylum acanthopodium Dc) terhadap Kultur Sel Melanoma B16F10

Abstract

Background: Melanoma is a type of skin cancer with increasing prevalence and a poor prognosis when lesions penetrate to dermis. Melanoma cells exhibit high proliferation rates and low responsiveness to available chemotherapy and treatments. The utilization of natural plant-derived substances is one option, and research on the potential of natural compounds as anticancer agents is ongoing. One such study focuses on the utilization of andaliman fruit extract (Zanthoxylum acanthopodium DC). Andaliman fruit extract is known to contain several compounds with potential anticancer properties, as previous studies have shown its effectiveness against breast and cervical cancer cells. Objectives: To investigate the effects of andaliman fruit extract on B16F10 melanoma cell cultures. The extract was obtained through maceration of its simplisia, followed by sequential fractionation using ethanol, n-hexane, ethyl acetate, and water solutions. Chemical compound identification and antioxidant testing were performed, continued by assay of cytotoxicity, inhibition of cell cycle progression using PI/RNase markers, inhibition of G1 phase and G1-S phase cell cycle using Cyclin D1 and Cyclin E, apoptosis induction using Annexin V and Caspase 3. These examinations were performed using flow cytometry. Results: The ethyl acetate fraction of andaliman fruit extract contains flavonoids, glycosides, triterpen, tannin, alkaloids, saponin, and steroid and had strong antioxidant activity. The cytotoxicity test found the IC50 value of 249.33 μg/ml and the treatment cells were grouped into 1IC50, ¾IC50 (187μg/ml), and ½IC50 (124.5 μg/ml). The 1IC50 and ¾IC50 inhibited the G1 phase, the 1IC50 and ½IC50 inhibited the S phase, and the 1IC50, ¾IC50, and ½IC50 inhibited the G2-M phase. The 1IC50, ¾IC50, and ½IC50 decreased the expression of Cyclin D1 and Cyclin E. The apoptosis induction assay using the Annexin V showed that the ¾IC50 increased apoptotic cells percentage not significantly, and the 1IC50 increased Caspase 3 expression significantly. This difference because the apoptotic cells had not yet exposed the phosphatidylserine in their outer membrane, so the Annexin V unable to bind with it. Conclusion: The ethyl acetate fraction of andaliman fruit extract exhibits potential as an anticancer agent by inhibiting cell cycle progression in the G1, S, and G2-M phases, and decreased the expression of Cyclin D1 and Cyclin E proteins, also demonstrates its ability to induce apoptosis through the increased of Caspase 3 expression.