Waziri Peter M, Abdullah Rasedee, Yeap Swee Keong, Omar Abdul Rahman, Abdul Ahmad Bustamam, Kassim Nur Kartinee, Malami Ibrahim, Karunakaran Thiruventhan, Imam Mustapha Umar
MAKNA Cancer Research Laboratory, Institute of Bioscience, University Putra Malaysia, Serdang, Selangor, Malaysia; Department of Biochemistry, Kaduna State University, Main Campus, PMB 2336 Kaduna, Nigeria.
Department of Veterinary Pathology and Microbiology, Faculty of Veterinary, University Putra Malaysia, Serdang, Selangor, Malaysia.
J Ethnopharmacol. 2016 Dec 24;194:549-558. doi: 10.1016/j.jep.2016.10.030. Epub 2016 Oct 8.
Clausena excavata Burm.f. is used locally in folk medicine for the treatment of cancer in South East Asia.
To determine the mechanism of action of pure clausenidin crystals in the induction of hepatocellular carcinoma (hepG2) cells apoptosis.
Pure clausenidin was isolated from Clausena excavata Burm.f. and characterized using H and C NMR spectra. Clausenidin-induced cytotoxicity was determined by MTT assay. The morphology of hepG2 after treatment with clausenidin was determined by fluorescence and Scanning Electron Microscopy. The effect of clausenidin on the apoptotic genes and proteins were determined by real-time qPCR and protein array profiling, respectively. The involvement of the mitochondria in clausenidin-induced apoptosis was investigated using MMP, caspase 3 and 9 assays.
Clausenidin induced significant (p<0.05) and dose-dependent apoptosis of hepG2 cells. Cell cycle assay showed that clausenidin induced a G2/M phase arrest, caused mitochondrial membrane depolarization and significantly (p<0.05) increased expression of caspases 3 and 9, which suggest the involvement of the mitochondria in the apoptotic signals. In addition, clausenidin caused decreased expression of the anti-apoptotic protein, Bcl 2 and increased expression of the pro-apoptotic protein, Bax. This finding was confirmed by the downregulation of Bcl-2 gene and upregulation of the Bax gene in the treated hepG2 cells.
Clausenidin extracted from Clausena excavata Burm.f. is an anti-hepG2 cell compound as shown by its ability to induce apoptosis through the mitochondrial pathway of apoptosis. Clausenidin can potentially be developed into an anticancer compound.
在东南亚，土蜜树在民间医学中被用于治疗癌症。
确定纯吴茱萸素晶体诱导肝癌（hepG2）细胞凋亡的作用机制。
从土蜜树中分离出纯吴茱萸素，并通过氢谱和碳谱核磁共振光谱进行表征。采用MTT法测定吴茱萸素诱导的细胞毒性。通过荧光显微镜和扫描电子显微镜观察吴茱萸素处理后hepG2细胞的形态。分别采用实时定量聚合酶链反应和蛋白质阵列分析测定吴茱萸素对凋亡基因和蛋白质的影响。使用线粒体膜电位、半胱天冬酶3和9检测法研究线粒体在吴茱萸素诱导的凋亡中的作用。
吴茱萸素诱导hepG2细胞发生显著（p<0.05）且剂量依赖性的凋亡。细胞周期分析表明，吴茱萸素诱导细胞阻滞于G2/M期，导致线粒体膜去极化，并显著（p<0.05）增加半胱天冬酶3和9的表达，这表明线粒体参与了凋亡信号传导。此外，吴茱萸素导致抗凋亡蛋白Bcl-2表达降低，促凋亡蛋白Bax表达增加。在经处理的hepG2细胞中，Bcl-2基因下调和Bax基因上调证实了这一发现。
从土蜜树中提取的吴茱萸素是一种抗hepG2细胞化合物，因其能够通过凋亡的线粒体途径诱导凋亡。吴茱萸素有可能被开发成为一种抗癌化合物。
