PDF(Pubmed)
Abstract：
Natural products contribute substantially to anticancer therapy; the plant kingdom provides an important source of molecules. Conofolidine is a novel Aspidosperma-Aspidosperma bisindole alkaloid isolated from the Malayan plant Tabernaemontana corymbosa. 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.
摘要：
天然产物对抗癌治疗有很大贡献；植物界提供了重要的分子来源。Conofolidine是一种新型的Aspidosperma-Aspidosperma双吲哚生物碱，分离自马来亚植物TabernaemontanaCorymbosa。在这里，我们报道了与其他三种双吲哚-康茶碱一起具有广谱抗癌活性，leuchyllidine,和双茶碱-针对人类乳腺，结直肠,胰腺,和肺癌细胞系。值得注意的是,conofolidine能够诱导细胞凋亡(例如，在MDA-MB-468乳房中)或衰老(例如，在HT-29结直肠中）在癌细胞中。膜联蛋白V-FITC/PI,caspase激活，PARP裂解证实了前者，而阳性β-gal染色证实了后者。细胞周期扰动是明显的，包括S相耗尽，伴随着CDK2和细胞周期蛋白（A2，D1）下调，p21上调。HCT-116细胞的共聚焦成像揭示了异常有丝分裂表型-膜起泡的诱导，DNA片段化与偶尔的多核化。在HCT-116，MDA-MB-468，MIAPaCa-2和HT-29细胞中的DNA完整性评估显示在G1细胞周期期间荧光γ-H2AX增加；通过共聚焦显微镜在HCT-116和MDA-MB-468细胞中验证了γ-H2AX灶。Conopolidine增加氧化应激，在大多数癌细胞系中，先前的凋亡和衰老诱导，如ROS水平增强伴随NQO1表达增加所见。总的来说，我们提出了一种推定的有效抗癌剂，能够在体外诱导异质模式的癌细胞死亡，鼓励对这种天然产品进行进一步的临床前评估。
