Eli Halimah
Gofarana Wilar
Ferry Ferdiansyah Sofian
Sandra Megantara
Jutti Levita

Introduction: Costus cuspidatus and Costus subsessilis have shown cytotoxic effects in HL۶۰, Jurkat, and THP-۱ cells. This study aimed to evaluate the cytotoxicity and apoptotic activity of the ethanol extract and fractions of C. speciosus leaves against MCF-۷ and HeLa cells and to study the binding affinity of known phytoconstituents in C. speciosus toward caspase-۳. Methods: The leaves were extracted with ۹۶% ethanol for ۷۲ hours. The extract was further subjected to a liquid-liquid fractionation by employing water, n-hexane, and ethyl acetate solvents. The cytotoxicity of the ethanol extract (CSEE), as well as water (CSWF), n-hexane (CSHF), and ethyl acetate (CSEF) fractions on MCF-۷ and HeLa cells was carried out using the CCK-۸/WST-۸ reagent, followed by flow cytometry analysis to obtain the apoptotic activity on both cells. Results: On MCF-۷ cells, the best cytotoxicity was exhibited by CSEF (IC۵۰ of ۵۸.۷۱ µg/mL), followed respectively by CSHF, CSWF, and CSEE. On HeLa cells, the extract and fractions weakly inhibited the survival growth rate, with the best cytotoxicity respectively shown by CSEF (IC۵۰ of ۲۳۳.۸۸۱ µg/mL), CSEE, and CSHF. Furthermore, the exposure of CSEF to MCF۷ cells resulted in an average of ۲۲.۹% undergoing early-stage apoptosis, to HeLa cells resulted in an average of ۴۰.۸۵% undergoing necrosis. Molecular docking simulation revealed that diosgenin and beta-sitosterol interacted with caspase-۳ residues with considerable affinity. Conclusion: The CSEF of C. speciosus leaves induces cytotoxicity, whose mechanism seems to be either at the early apoptosis stage on MCF-۷ cells or by necrosis on HeLa cells.