依地酮和依地酯是长叶肉苁蓉的根和茎中已知的苦参内酯类化合物。这些化合物已被报道具有细胞毒性效应，然而其在几种癌细胞系中的作用机制尚未阐明。本研究旨在通过硫苏胺B（Sulforhodamine B）测定法，研究依地酮和依地酯在宫颈癌（HeLa）、结直肠癌（HT29）和卵巢癌（A2780）细胞系中的抗癌效应和作用机制。其细胞死亡机制是基于Hoechst 33342测定法和体外分子对接模拟以确定丁酸还原酶（DHFR）和肿瘤坏死因子α（TNF-α）是否为可能的蛋白靶点。依地酮和依地酯表现出体外抗癌效应，其IC50值分别为4.58 ± 0.090 µM 和1.60 ± 0.12 µM（HeLa）、1.22 ± 0.11 µM 和2.21 ± 0.049 µM（HT-29）、以及1.37 ± 0.13 µM 和2.46 ± 0.081 µM（A2780）。它们以剂量和时间依赖的方式诱导细胞凋亡。根据蛋白对接模拟结果，依地酮和依地酯显示出良好的抑制潜力，与TNF-α结合的结合亲和力为-8.83和-7.51kcal/mol，与DHFR结合的结合亲和力分别为-8.05和-8.87 kcal/mol。这些结果支持了依地酮和依地酯作为通过细胞凋亡机制的抗癌药物的证据，其与TNF-α和DHFR的抑制可能与之相关。

Eurycomanone and eurycomalactone are known quassinoids present in the roots and stems of Eurycoma longifolia. These compounds had been reported to have cytotoxic effects, however, their mechanism of action in a few cancer cell lines have yet to be elucidated. This study was aimed at investigating the anticancer effects and mechanisms of action of eurycomanone and eurycomalactone in cervical (HeLa), colorectal (HT29) and ovarian (A2780) cancer cell lines via Sulforhodamine B assay. Their mechanism of cell death was evaluated based on Hoechst 33342 assay and in silico molecular docking toward DHFR and TNF-α as putative protein targets. Eurycomanone and eurycomalactone exhibited in vitro anticancer effects manifesting IC50 values of 4.58 ± 0.090 µM and 1.60 ± 0.12 µM (HeLa), 1.22 ± 0.11 µM and 2.21 ± 0.049 µM (HT-29), and 1.37 ± 0.13 µM and 2.46 ± 0.081 µM (A2780), respectively. They induced apoptotic cancer cell death in dose- and time-dependent manners. Both eurycomanone and eurycomalactone were also predicted to have good inhibitory potential as demonstrated by the docking into TNF-α with binding affinity of -8.83 and -7.51 kcal/mol, respectively, as well as into DHFR with binding affinity results of -8.05 and -8.87 kcal/mol, respectively. These results support the evidence of eurycomanone and eurycomalactone as anticancer agents via apoptotic cell death mechanism that could be associated with TNF-α and DHFR inhibition as among possible protein targets.