Department of Pharmacy, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
Evid Based Complement Alternat Med. 2013;2013:689865. doi: 10.1155/2013/689865. Epub 2013 Mar 13.
Murraya koenigii Spreng has been traditionally claimed as a remedy for cancer. The current study investigated the anticancer effects of girinimbine, a carbazole alkaloid isolated from Murraya koenigii Spreng, on A549 lung cancer cells in relation to apoptotic mechanistic pathway. Girinimbine was isolated from Murraya koenigii Spreng. The antiproliferative activity was assayed using MTT and the apoptosis detection was done by annexin V and lysosomal stability assays. Multiparameter cytotoxicity assays were performed to investigate the change in mitochondrial membrane potential and cytochrome c translocation. ROS, caspase, and human apoptosis proteome profiler assays were done to investigate the apoptotic mechanism of cell death. The MTT assay revealed that the girinimbine induces cell death with an IC50 of 19.01 μ M. A significant induction of early phase of apoptosis was shown by annexin V and lysosomal stability assays. After 24 h treatment with 19.01 μ M of girinimbine, decrease in the nuclear area and increase in mitochondrial membrane potential and plasma membrane permeability were readily visible. Moreover the translocation of cytochrome c also was observed. Girinimbine mediates its antiproliferative and apoptotic effects through up- and downregulation of apoptotic and antiapoptotic proteins. There was a significant involvement of both intrinsic and extrinsic pathways. Moreover, the upregulation of p53 as well as the cell proliferation repressor proteins, p27 and p21, and the significant role of insulin/IGF-1 signaling were also identified. Moreover the caspases 3 and 8 were found to be significantly activated. Our results taken together indicated that girinimbine may be a potential agent for anticancer drug development.
九里香已被传统上用于治疗癌症。本研究调查了从九里香中分离得到的咔唑生物碱吉尼定对 A549 肺癌细胞的抗癌作用，以及与凋亡机制途径的关系。吉尼定是从九里香中分离出来的。采用 MTT 法测定增殖活性，用 Annexin V 和溶酶体稳定性测定法检测凋亡。进行多参数细胞毒性测定以研究线粒体膜电位和细胞色素 c 易位的变化。进行 ROS、caspase 和人类凋亡蛋白谱分析以研究细胞死亡的凋亡机制。MTT 法显示吉尼定诱导细胞死亡的 IC50 为 19.01 μM。 Annexin V 和溶酶体稳定性测定显示早期凋亡明显诱导。用 19.01 μM 的吉尼定处理 24 小时后，很容易观察到核区缩小、线粒体膜电位增加和质膜通透性增加。还观察到细胞色素 c 的易位。吉尼定通过上调和下调凋亡和抗凋亡蛋白来介导其增殖抑制和凋亡作用。涉及到内在和外在途径。此外，还鉴定了 p53 的上调以及细胞增殖抑制剂蛋白 p27 和 p21 的显著作用，以及胰岛素/IGF-1 信号的重要作用。还发现 caspase 3 和 8 被显著激活。我们的综合结果表明，吉尼定可能是抗癌药物开发的潜在药物。
