RESUMO
In the present study, A fluorescent imaging-based high-throughput screening method was developed for identifying anti-migratory compounds with 96-well Transwell plates. The correlation, precision and stability of this method were examined and the incubation time of dye Hoechst 33342 in addition to migration time was optimized. In addition, The inhibitory activity of anti-cancer drug paclitaxel on tumor cell migration was assayed and an IC50 value of 0.717 micromol x L(-1) was obtained. Using this method, 24 components from Rhizoma Alismatis were screened and one component with anti-migration activity was found. These results show that the new proposed method with good precision, stability and linear range has the potential to assay the inhibitory activity of anticancer compounds.
Assuntos
Humanos , Alisma , Química , Antineoplásicos Fitogênicos , Farmacologia , Linhagem Celular Tumoral , Movimento Celular , Ensaios de Seleção de Medicamentos Antitumorais , Métodos , Medicamentos de Ervas Chinesas , Farmacologia , Fluorescência , Ensaios de Triagem em Larga Escala , Métodos , Concentração Inibidora 50 , Paclitaxel , Farmacologia , Plantas Medicinais , Química , Rizoma , QuímicaRESUMO
<p><b>OBJECTIVE</b>To develop a fluorescence imaging-based novel system for quick screening of antitumor compounds in vitro.</p><p><b>METHODS</b>The antitumor activity of 26 components from Lindera aggregate were determined by relative number of viable cell labelled with fluorescein diacetate (FDA) in multiwell plates after exposure to these 26 different components. Then, the linearity and precision of this method were validated. The structures of active compounds in components with strong antitumor activity were deduced by LC/MS.</p><p><b>RESULTS</b>The linearity of this method for cells stained with FDA was validated (r² = 0.9858) in the range of 0-10⁴ cells per well, and the in-plate precision was 9.41 %. Two of 26 components from Lindera aggregate showed significant inhibition effect on proliferation of HepG2 cells (inhibition rate >90%).</p><p><b>CONCLUSION</b>This proposed rapid and reliable approach can be used for screening compounds with antitumor activity from Traditional Chinese Medicine in vitro. The major active compound of Lindera aggregate was putatively identified as norboldine by LC/MS analysis.</p>