Single-Dose and Combined-Dose of Nanoparticles from Soursop Leaves (Annona muricata L.) and Sappan Wood (Caesalpinia sappan L.) Induced Apoptosis and Necrosis in HeLA Cells

Introduction: Apart from the medical advancement of chemotherapy, various plants were known as beneficial for cancer therapy because they can kill cancer cells selectively without damaging the normal cells. Here, we showed that nanoparticles formulated from chloroform fraction of soursop (Annona muricata L.) leaves and ethyl acetate fraction of sappan wood (Caesalpinia sappan L.) have anti-proliferative and cytotoxic effects on HeLa cervical cancer cells. Methods: The cytotoxic effect was evaluated using a single dose of each nanoparticle and a combined dose to obtain a synergistic effect. The mechanism of induced cell death via apoptosis or necrosis pathway was evaluated using flow cytometry by incorporating Annexin V and propidium iodide. Results: Synthesis of nanoparticles from the extract of soursop leaves (nano-SL) and extract of sappan wood (nano-SW) yielded particle sizes ranging from 248 to 317 nm. Nano-SL and nano-SW decreased the viability of HeLa cervical cancer cells in a dose-dependent manner with IC₅₀ values of 63,32 μg/ml dan 40,88 μg/ml, respectively. The combined dose of 1/8 IC₅₀ from both nanoparticles showed a strong synergistic effect, as shown by the combination index value of 0.13 based on the same mode of action and different modes of action. In HeLa cells treated with a combined dose of nanoparticles, the total apoptotic cells increased two times greater than that in control cells. Conclusion: Nano-SL and nano-SW induce apoptosis and necrosis in HeLa cells. Combined-dose of both nanoparticles produced a synergistic effect that could reduce the amount of the required individual dose while increasing the total effect.