rosmarinic-acid and Precursor-Cell-Lymphoblastic-Leukemia-Lymphoma

Salvia miltiorrhiza (SM) Bunge is one of the widely-used Chinese medicinal herbs. In this study, the chemical constituents and anticancer potential of SM stems and leaves were examined with those of respective callus cultures. The callus culture for stem and leaf explants was initiated in modified Murashige and Skoog (MS) medium. Active constituents of respective extracts were analyzed by high performance liquid chromatography coupled with DAD and MS (HPLC-DAD-MS). Rosmarinic acid (RA) and salvianolic acid B (Sal B) were determined to be the main phenolic compounds. Quantitative analyses revealed that callus stem extracts produced higher amount of RA and Sal B (stem RA: 1.27±0.38%; stem Sal B: 0.87±0.20%) than callus leaf did (leaf RA: 0.28±0.02%; leaf Sal B: 0.07±0.03%). Stem and leaf callus extracts exerted cytotoxic effects towards CCRF-CEM cells (stem: 13.1±0.90 μg/ml; leaf: 18.1±0.33 μg/ml). As expected, stem extract with higher amount of RA and Sal B showed lower IC50 value than leaf extract. These findings suggest the possibility to isolate bioactive constituents with anticancer properties from in vitro callus cultures of stems and leaves of SM.

Topics: Benzofurans; Chromatography, High Pressure Liquid; Cinnamates; Depsides; Drugs, Chinese Herbal; Plant Extracts; Plant Leaves; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Rosmarinic Acid; Salvia miltiorrhiza

Rosmarinic acid (RA), a major hydrosoluble bioactive compound found in the Chinese medicinal herb, Salvia miltiorrhiza Bunge, which has been used in traditional Chinese medicine to treat various diseases, including cancer. However, the mechanisms have not been fully elucidated.. Guided by microarray hybridization and Ingenuity Pathway Analysis, we identified modes of action of rosmarinic acid (RA) isolated from S. miltiorrhiza on acute lymphoblastic leukemia cells.. Microarray data were verified by independent methods: Real-time RT-PCR (mRNA expression), resazurin assay (cytotoxicity of RA towards parental CCRF-CEM, multidrug-resistant CEM/ADR5000 cells and normal lymphocytes), flow cytometry (cell cycle arrest, apoptosis, necroptosis, generation of reactive oxygen species (ROS), disruption of mitochondrial membrane potential (MMP)), single cell gel electrophoresis (DNA damage), molecular docking and gene promoter binding motif analysis (NFκB), Western blotting (nuclear NFκB translocation, PARP cleavage, caspase 3/7/9 expression), and fibronectin-based cell adhesion assay.. RA dose-dependently inhibited CCRF-CEM and CEM/ADR5000 cells, but caused less cytotoxicity towards normal lymphocytes. RA simultaneously induced apoptosis and necrosis, as shown by cell morphology and annexin V-PI assay. DNA damage was dose-dependently induced without ROS generation, which subsequently led to cell cycle arrest. RA-stimulated MMP dysfunction activated PARP-cleavage and caspase-independent apoptosis. In accordance with molecular docking and gene promoter binding motif analyses, p65 translocation from the cytosol to the nucleus was blocked by RA, indicating a mechanistic role of the NFκB pathway to explain RA's action. RA affected cellular movement as evaluated by ameliorating cell adhesion to fibronectin.. RA induced apoptosis and necrosis in a ROS-independent DNA damage and caspase-independent manner. These results may contribute to the rationale use of S. miltiorrhiza and RA in traditional medicine of leukemia.

Topics: Antineoplastic Agents; Apoptosis; Cell Adhesion; Cell Cycle; Cell Line, Tumor; Cell Survival; Cells, Cultured; Cinnamates; Comet Assay; Depsides; DNA Damage; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Gene Expression Profiling; Humans; Lymphocytes; Membrane Potential, Mitochondrial; Molecular Docking Simulation; NF-kappa B; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Reactive Oxygen Species; Rosmarinic Acid; Salvia miltiorrhiza