School of Pharmacy, Al-Kitab University, Kirkuk 36015, Iraq.
School of Pharmacy, Biodiscovery Institute, University of Nottingham, University Park, Nottingham NG7 2RD, UK.
Molecules. 2024 Jun 4;29(11):2654. doi: 10.3390/molecules29112654.
Natural products contribute substantially to anticancer therapy; the plant kingdom provides an important source of molecules. Conofolidine is a novel bisindole alkaloid isolated from the Malayan plant . Herein, we report conofolidine's broad-spectrum anticancer activity together with that of three other bisindoles-conophylline, leucophyllidine, and bipleiophylline-against human-derived breast, colorectal, pancreatic, and lung carcinoma cell lines. Remarkably, conofolidine was able to induce apoptosis (e.g., in MDA-MB-468 breast) or senescence (e.g., in HT-29 colorectal) in cancer cells. Annexin V-FITC/PI, caspase activation, and PARP cleavage confirmed the former while positive β-gal staining corroborated the latter. Cell cycle perturbations were evident, comprising S-phase depletion, accompanied by downregulated CDK2, and cyclins (A2, D1) with p21 upregulation. Confocal imaging of HCT-116 cells revealed an induction of aberrant mitotic phenotypes-membrane blebbing, DNA-fragmentation with occasional multi-nucleation. DNA integrity assessment in HCT-116, MDA-MB-468, MIAPaCa-2, and HT-29 cells showed increased fluorescent γ-H2AX during the G1 cell cycle phase; γ-H2AX foci were validated in HCT-116 and MDA-MB-468 cells by confocal microscopy. Conofolidine increased oxidative stress, preceding apoptosis- and senescence-induction in most carcinoma cell lines as seen by enhanced ROS levels accompanied by increased NQO1 expression. Collectively, we present conofolidine as a putative potent anticancer agent capable of inducing heterogeneous modes of cancerous cell death in vitro, encouraging further preclinical evaluations of this natural product.
天然产物在癌症治疗中做出了重要贡献；植物王国是分子的重要来源。从马来植物中分离得到的新型双吲哚生物碱——苦参定具有广谱抗癌活性。此外，本文还报道了三种其他双吲哚类化合物——汉防己甲素、白屈菜丙素和比枯枯灵——与苦参定一起对人源性乳腺癌、结直肠癌、胰腺癌和肺癌细胞系的抗癌活性。值得注意的是，苦参定能够诱导癌细胞凋亡（如 MDA-MB-468 乳腺癌）或衰老（如 HT-29 结直肠癌细胞）。 Annexin V-FITC/PI、半胱天冬酶激活和 PARP 切割证实了前者，而β-半乳糖苷酶染色阳性则证实了后者。细胞周期受到明显干扰，包括 S 期耗竭，同时伴随着 CDK2 和细胞周期蛋白（A2、D1）下调以及 p21 上调。HCT-116 细胞的共聚焦成像显示诱导了异常有丝分裂表型——细胞膜起泡、DNA 碎片化，偶尔出现多核化。HCT-116、MDA-MB-468、MIAPaCa-2 和 HT-29 细胞的 DNA 完整性评估显示，在 G1 细胞周期阶段荧光 γ-H2AX 增加；通过共聚焦显微镜验证了 HCT-116 和 MDA-MB-468 细胞中的 γ-H2AX 焦点。苦参定在大多数癌细胞系中增加氧化应激，导致凋亡和衰老诱导，表现为 ROS 水平升高，同时 NQO1 表达增加。综上所述，我们提出苦参定是一种有潜力的抗癌药物，能够在体外诱导多种癌细胞死亡模式，这鼓励进一步对该天然产物进行临床前评估。
