Inhibition of the RhoA GTPase Activity Increases Sensitivity of Melanoma Cells to UV Radiation Effects
Scratch wound healing and Matrigel invasion assays for MeWo, MeWo-RhoA-N19, and MeWo-RhoA-V14 cells treated with UV radiation. (a) A scratch-like wound was made in a monolayer of cells on 100% confluent plates using a micropipette tip (time zero) prior to irradiation treatment (50 KJ/m2 UVA, 80 J/m2 UVB, or 4 J/m2 UVC). The cells were photographed at 0 and 24 h after treatment at 20x magnification using an inverted microscope (Olympus, Tokyo, Japan), and representative micrographs are shown. (b) Measurements of the initial and final open areas were calculated using cell-F software (Olympus, Tokyo, Japan) and were plotted in bar graphs as the percentage of the closed area. The results are presented as the mean and standard deviation from at least three independent images captured at 24 h after treatment. Two-way ANOVA was performed to compare the RhoA-N19 clone with the two other clones treated according to the same specified conditions. . (c) Representative micrographs of the Matrigel invasion assay for MeWo, MeWo-RhoA-N19, and MeWo-RhoA-V14 cells untreated (control) or pretreated with 4 J/m2 UVC or with 25 μM of a broad-spectrum MMP inhibitor (GM6001). Nuclei of the invasive cells were visualized using DAPI (4x magnification). (d) Quantification of invasive cells shown in 2C. A -test was performed to compare the control cells with the UVC radiation-treated cells from two independent experiments. ; ns, nonsignificant.