G1 arrest and caspase-mediated apoptosis in HL-60 cells by dichloromethane extract of Centrosema pubescens.
Cell division and apoptosis are two crucial components of tumor biology and the importance of increased cell proliferation and reduced cell death have made them valid therapeutic targets. The plant kingdom is a relatively underexploited cache of novel drugs, and crude extracts of plants are known for their synergistic activity. The present study assessed the anti-proliferative activity of the medicinal plant Centrosema pubescens Benth. Centrosema pubescens dichloromethane extract (CPDE) inhibited the proliferation of HL-60 (promyelocytic acute leukaemia) cells with an IC₅₀ value of 5 μg/ml. Further studies also showed that CPDE induces growth arrest at the G1 phase and specifically down-regulates the expressions of cyclin E and CDK2 and up-regulates p27(CKI) levels. These events apparently lead to the induction of apoptosis, which was demonstrated qualitatively by a DNA fragmentation assay and propidium iodide staining. Quantitative assessment of the effective arrest of the cell cycle and of apoptosis was confirmed by flow cytometry. CPDE exhibited negligible cytotoxicity even at the highest dose tested (100 μg/ml) in both normal peripheral blood mononuclear cells and in an in vitro model (HL-60). Our results strongly suggest that CPDE arrests the cell cycle at the G1 phase and triggers apoptosis by caspase activation.
Centre for Biotechnology, Anna University, Chennai, India.
Subramanireddy
S
Ramadevi Mani
Baddireddi Subhadra
BS
Lakshmi
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D019927:Cyclin E / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D053938:DNA Fragmentation; D007887:Fabaceae; D016193:G1 Phase / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy* / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology* / Q000627:therapeutic use; D018515:Plant Leaves
D000972:Antineoplastic Agents, Phytogenic; D019927:Cyclin E; D010936:Plant Extracts; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D020169:Caspases