This study was aimed to investigate the cytotoxic potential of a natural compound, progenin III on a broad range of cancer cell lines, including various sensitive and drug-resistant phenotypes. The cytotoxicity, progenin III-induced autophagic, ferroptotic and necroptotic cell death were evaluated by the resazurin reduction assay (RRA). Spectrophotometric analysis of caspases activity was performed using caspase-Glo assay. Flow cytometry was applied for cell cycle analysis (PI staining), apoptosis (annexin V/PI staining), mitochondrial membrane potential (MMP) (JC-1) and reactive oxygen species (ROS) (H₂DCFH-DA). Progenin III and the reference molecule, doxorubicin exerted cytotoxic effects towards the 18 cancer cell lines tested including animal and human cell lines. The IC₅₀ values obtained ranged from 1.59 μM (towards CCRF-CEM leukemia cells) to 31.61 μM (against the BRAF-V600E homozygous mutant SKMel-28 melanoma cells) for progenin III. Normal sensitivity was achieved with CEM/ADR5000 cells and HCT116p53^−/− adenocarcinoma cells respectively compared to their sensitive congeners CCRF-CEM cells and HCT116 p53^+/+ cells. Progenin III induced apoptosis in CCRF-CEM cells mediated by caspases 3/7 activation, MMP alteration and increase ROS production, and otherwise autophagy and necroptosis. Progenin III is a potential anticancer molecule that deserves further investigations to develop a novel drug to combat malignant diseases including refractory cancers.

这项研究旨在研究天然化合物Progenin III对多种癌细胞系（包括各种敏感和耐药表型）的细胞毒性潜力。通过刃天青素还原测定法（RRA）评估了细胞毒性，progenin III诱导的自噬，铁促和坏死性细胞死亡。使用胱天蛋白酶-Glo测定法对胱天蛋白酶活性进行分光光度分析。流式细胞仪用于细胞周期分析（PI染色），凋亡（annexin V / PI染色），线粒体膜电位（MMP）（JC-1）和活性氧（ROS）（H ₂ DCFH-DA）。Progenin III和参考分子阿霉素对包括动物和人细胞系在内的18种癌细胞系产生了细胞毒性作用。IC ₅₀对于前体素III，获得的值范围从1.59μM（朝向CCRF-CEM白血病细胞）到31.61μM（针对BRAF-V600E纯合突变SKMel-28黑色素瘤细胞）。与它们的敏感同类CCRF-CEM细胞和HCT116 p53 ^{+ / +}细胞相比，分别使用CEM / ADR5000细胞和HCT116 p53 ^-/-腺癌细胞可达到正常敏感性。Progenin III诱导了胱天蛋白酶3/7激活，MMP改变和ROS产生以及自噬和坏死性坏死介导的CCRF-CEM细胞凋亡。Progenin III是一种潜在的抗癌分子，值得进一步研究以开发出一种新型药物来对抗包括难治性癌症在内的恶性疾病。