Cytotoxicity of epunctanone and four other phytochemicals isolated from the medicinal plants Garcinia epunctata and Ptycholobium contortum towards multi-factorial drug resistant cancer cells

Abstract Introduction Resistance of cancer cells is a serious impediment to chemotherapy and several phytochemicals are active against multi-drug resistant (MDR) phenotypes. The cytotoxicity of five naturally occurring compounds: betulin ( 1 ), mundulea lactone ( 2 ), seputhecarpan A ( 3 ), seputheisoflavone ( 4 ) and epunctanone ( 5 ) was evaluated on a panel of 9 cancer cell lines including various sensitive and drug-resistant cell lines. The modes of action of compound 5 were further investigated. Methods The resazurin reduction assay was used to evaluate cytotoxicity of samples and ferroptotic cell death induced by compound 5 ; caspase-Glo assay was used to detect the activation of caspases in CCRF-CEM leukemia cells treated with compound 5 . Flow cytometry was used for cell cycle analysis in CCRF-CEM cells treated with compound 5 , as well as detection of apoptotic cells by annexin V/PI staining, analysis of mitochondrial membrane potential (MMP) and measurement of reactive oxygen species (ROS). Results Compounds 1–5 displayed cytotoxic effects in the 9 studied cancer cell lines with IC 50 values below 70 µM. The IC 50 values varied from 8.20 µM (in HCT116 ( p53 −/− ) colon cancer cells) to 35.10 µM (against HepG2 hepatocarcinoma cells) for 1 , from 8.84 µM (in CEM/ADR5000 leukemia cells) to 48.99 µM (in MDA-MB-231 breast adenocarcinoma cells) for 2 , from 12.17 µM (in CEM/ADR5000 cells) to 65.08 µM (in MDA-MB-231 cells) for 3 , from 23.80 µM (in U87MG.Δ EGFR glioblastoma cells) to 68.66 µM (in HCT116 ( p53 −/− ) cells) for 4 , from 4.84 µM (in HCT116 ( p53 −/− ) cells) to 13.12 µM (in HepG2 cells) for 5 and from 0.02 µM (against CCRF-CEM cells) to 122.96 µM (in CEM/ADR5000 cells) for doxorubicin. Compound 5 induced apoptosis in CCRF-CEM cells through alteration of MMP and increase in ROS production. In addition to apoptosis, ferroptosis was also identified as another mode of cell death induced by epunctanone. Conclusions Compounds 1–5 are valuable cytotoxic compounds that could be used to combat MDR cancer cells. Benzophenoe 5 is the most active molecule and deserve more investigations to develop new anticancer drugs.