title: Effects of 5,6-Dihydroxy-2,4-Dimethoxy-9,10-Dihydrophenanthrene on G2/M Cell Cycle Arrest and Apoptosis in Human Lung Carcinoma Cells.
abstract: 5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene (HMP) is an active compound isolated from the rhizome extracts of Dioscorea membranacea Pierre, a Thai medicinal plant. This study aimed to investigate the growth-inhibitory and apoptosis-inducing effects of HMP in human lung cancer A549 cells. The antiproliferative and cytotoxic effects of HMP were analyzed by a Sulforhodamine B assay. Cell division, cell cycle distribution and membrane asymmetry changes were each performed with different fluorescent dyes and then analyzed by flow cytometry. Real-time PCR and immunoblotting were used to detect cell cycle- and apoptosis-related mRNA levels and proteins, respectively. The nuclear morphology of the cells stained with DAPI and DNA fragmentation were detected by fluorescence microscopy and gel electrophoresis, respectively. The results showed that HMP exerted strong antiproliferative and cytotoxic activities in A549 cells with the highest selectivity index. It halted the cell cycle in [Formula: see text]/M phase via down-regulation of the expression levels of regulatory proteins Cdc25C, Cdk1 and cyclinB1. In addition, HMP induced early apoptotic cells with externalized phosphatidylserine and subsequent apoptotic cells in sub-[Formula: see text] phase. HMP increased caspase-3 activity and levels of the cleaved (active) form of caspase-3 whose actions were supported by the cleavage of its target PARP, nuclear condensation and DNA apoptotic ladder. Moreover, HMP significantly increased the mRNA and protein levels of proapoptotic Bax as well as promoted subsequent caspase-9 activation and BID cleavage, indicating HMP-induced apoptosis via both intrinsic and extrinsic pathways. These data support, for the first time, the potential role of HMP as a cell-cycle arrest and apoptosis-inducing agent for lung cancer treatment.
authors: * Biochemistry and Molecular Biology, Department of Preclinical Science, Faculty of Medicine, Thailand.
authors: Wipada
authors: W
authors: Duangprompo
authors: † Microbiology and Immunology, Department of Preclinical Science, Faculty of Medicine, Thailand.
authors: Kalaya
authors: K
authors: Aree
authors: ‡ Department of Applied Thai Traditional Medicine, Faculty of Medicine, Thailand.
authors: Arunporn
authors: A
authors: Itharat
authors: § Biochemistry, Department of Preclinical Science, Faculty of Medicine, Thammasat University (Rangsit Campus), Klongluang, Thailand.
authors: Pintusorn
authors: P
authors: Hansakul
mesh_terms: D000072283:A549 Cells; D000972:Antineoplastic Agents, Phytogenic; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000473:pathology*; D020169:Caspases / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects* / Q000235:genetics; D027742:Dioscorea / Q000737:chemistry*; D015536:Down-Regulation / Q000187:drug effects; D016195:G2 Phase / Q000187:drug effects* / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy / Q000473:pathology*; D010616:Phenanthrenes / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D008517:Phytotherapy; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: C000613699:5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene; D000972:Antineoplastic Agents, Phytogenic; D018797:Cell Cycle Proteins; D010616:Phenanthrenes; D010936:Plant Extracts; D020169:Caspases
keywords: 5,6-Dihydroxy-2,4-Dimethoxy-9,10-Dihydrophenanthrene; Anticancer Effect; Apoptosis; Cell Cycle Arrest; Dioscorea membranacea Pierre