Dihydromyricetin inhibits migration and invasion of hepatoma cells through regulation of MMP-9 expression

Figure 4 Dihydromyricetin inhibits cell invasion and adhesion in vitro. Cell invasion assays were performed using a method similar to the cell motility assays, but using 8.0-μm pore size polycarbonate membranes that were pre-coated with 100 μL of Matrigel and allowed to solidify in an incubator at 37 °C for 3-4 h as described in the ‘‘MATERIALS AND METHODS’’. The SK-Hep-1 (A) and MHCC97L (C) cells that passed through the Matrigel and membrane were fixed in 75% ethanol and stained with hematoxylin and eosin solution. The number of SK-Hep-1 (B) and MHCC97L (D) cells that passed through Matrigel and membrane was quantified by determining the number of cells per field under an inverted microscope at 100 × magnification. The cells in at least four fields of each bottom of chamber were randomly photographed and counted for each assay. DHM treatment significantly inhibited the invasion of SK-Hep-1 and MHCC97L cells. Each experiment was performed at least three times. To measure the effect of DHM treatment on cell adhesion, SK-Hep-1 and MHCC97L cells were pre-treated with or without DHM at the indicated concentrations for 24 h. The cells were then resuspended in serum-free culture medium and added to 96-well plates coated with fibronectin. After incubation for 1 h, the adherent cells were measured using MTT assays. Dihydromyricetin (DHM) treatment decreased the number of SK-Hep-1 (E) and MHCC97L (F) cells adhered to fibronectin compared to the control. ^bP < 0.01 vs control, Student’s t test.