ABSTRACT
BACKGROUND: Isofraxidin (IF), a natural coumarin compound, has been reported to possess anti-cancer activity in human liver cancer. However, whether IF is involved in the regulation of colorectal cancer tumorigenesis and development has been not well elucidated. METHODS: The cell proliferation were assessed by Cell Counting Kit-8 (CCK-8) and colony formation test, respectively. The transwell assays were conducted to estimate cell migration and invasion abilities. Further, cell apoptosis was evaluated by confocal microscopy analysis, flow cytometry detection and TdT-mediated dUTP Nick-End Labeling (TUNEL) method. Western blot were performed to detect the expression of related protein. RESULTS: Herein, the result indicated that IF remarkably bated cell proliferation in human colorectal cancer cells HT-29 and SW-480 in a dose- and time-dependent manner. In addition, IF treatment showed obvious inhibitory activity to cell colony formation in HT-29 and SW-480 cells. Confocal microscopy analysis and flow cytometry detection revealed that IF dramatically induced cell apoptosis in HT-29 and SW-480 cells compared with the control. And IF markedly decreased the expression of anti-apoptotic protein bcl-2, whereas the expression of pro-apoptotic proteins, including caspase-3, caspase-9 and bax, notably increased in HT-29 and SW-480 cells. Besides, IF blocked Akt pathway via inhibition expression of p-Akt. Furthermore, MK2206, an Akt inhibitor, could inhibit cell colony formation and induced apoptosis. This effect is even more obvious in the presence of MK2206 and IF compared to that of either agent alone. CONCLUSIONS: Together, the present study reports a novel use of IF in mitigating human colorectal cancer proliferation and inducing apoptosis via blockage of Akt pathway.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Colorectal Neoplasms/drug therapy , Coumarins/pharmacology , Neoplasm Proteins/antagonists & inhibitors , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Signal Transduction , Antineoplastic Agents, Phytogenic/agonists , Caspase 3/chemistry , Caspase 3/metabolism , Caspase 9/chemistry , Caspase 9/metabolism , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/pathology , Coumarins/agonists , Drug Synergism , Heterocyclic Compounds, 3-Ring/pharmacology , Humans , Neoplasm Invasiveness/prevention & control , Neoplasm Proteins/agonists , Neoplasm Proteins/metabolism , Neoplastic Stem Cells/drug effects , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , Phosphorylation/drug effects , Protein Kinase Inhibitors/pharmacology , Protein Processing, Post-Translational/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-bcl-2/antagonists & inhibitors , Proto-Oncogene Proteins c-bcl-2/metabolism , bcl-2-Associated X Protein/agonists , bcl-2-Associated X Protein/metabolismABSTRACT
The present study investigated the antioxidant signalling mechanism of a coumarin-derived schiff base (CSB) scaffold against tert-butylhydroperoxide (TBHP) induced oxidative insult in murine hepatocytes. CSB possesses DPPH and other free radical scavenging activities. TBHP reduced cell viability and intracellular antioxidant status accompanied by an increase in intracellular ROS production in hepatocytes. TBHP also activated phospho-ERK1/2, phospho-p38 and NF-κB, altered the Bcl-2/Bad ratio, reduced mitochondrial membrane potential, released cytochrome C and activated caspase 3, suggesting that TBHP induced oxidative stress responsive cell death via apoptotic pathway. FACS analysis and DNA fragmentation studies also confirmed the apoptotic cell death in TBHP exposed hepatocytes. Treatment with CSB effectively reduced these adverse effects by preventing the oxidative insult, alteration in the redox-sensitive signalling cascades and mitochondrial events. Combining, results suggest that antioxidant property of CSB make the molecule to be a potential protective measure against oxidative insult, cytotoxicity and cell death.
Subject(s)
Antioxidants/pharmacology , Cell Death/drug effects , Coumarins/pharmacology , Cytoprotection , Membrane Potential, Mitochondrial/drug effects , Schiff Bases/pharmacology , tert-Butylhydroperoxide/toxicity , Animals , Apoptosis/drug effects , Caspase 3/drug effects , Caspase 3/metabolism , Cell Survival/drug effects , Coumarins/agonists , Coumarins/chemical synthesis , DNA Fragmentation/drug effects , Extracellular Signal-Regulated MAP Kinases/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , Free Radical Scavengers/pharmacology , Free Radicals/metabolism , Hepatocytes/drug effects , Lipid Peroxidation/drug effects , Mice , NF-kappa B/drug effects , NF-kappa B/metabolism , Oxidation-Reduction/drug effects , Oxidative Stress/drug effects , Oxidative Stress/physiology , Reactive Oxygen Species/metabolism , Reactive Oxygen Species/pharmacology , Schiff Bases/chemical synthesisABSTRACT
The responses of anthocyanin-producing (violet) and non-producing (white) cells of Glehnia littoralis to radical generators were compared. Cell growth, anthocyanin content, phenylalanine ammonia-lyase (PAL) activity and furanocoumarin production were determined after treatment with H(2)O(2), 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), X-ray and yeast extract, independently. AAPH and H(2)O(2) repressed the growth of both violet and white cells, but violet cells grew better than white cells. On the other hand, the anthocyanin content in violet cells decreased. Neither X-ray nor yeast extract affected cell growth or pigment production. Treatment with H(2)O(2), yeast extract, and X-ray, but not AAPH, induced PAL activity and furanocoumarin production in white cell cultures, whereas violet cell cultures did not produce furanocoumarin following any of the treatment employed.
Subject(s)
Amidines/pharmacology , Anthocyanins/metabolism , Apiaceae/drug effects , Hydrogen Peroxide/pharmacology , Phenylalanine Ammonia-Lyase/metabolism , Anthocyanins/radiation effects , Apiaceae/cytology , Apiaceae/metabolism , Cell Division/drug effects , Cell Division/radiation effects , Cell Line , Coumarins/agonists , Coumarins/metabolism , Coumarins/radiation effects , Phenylalanine Ammonia-Lyase/drug effects , Phenylalanine Ammonia-Lyase/radiation effects , X-RaysABSTRACT
Despite approximately 40 years of experience with oral anticoagulant drugs, controversy still exists about the safety of dental treatment in a patient receiving this therapy. The authors review the topic in depth and offer detailed recommendations for the dental management of patients receiving coumarin anticoagulant therapy.
