Figure 1.
Radiosensitivity of parental and established radioresistant sublines.
(A) Clonogenic cell survival curve. UPCI:SCC029B parental cells, 50Gy-UPCI:SCC029B intermediate radioresistant cells and 70Gy-UPCI:SCC029B final radioresistant cells. Data is represented as percentage survival of cells with mean (±SD) of three independent experiments. One-way ANOVA statistical analysis was performed on surviving fractions at the given doses, p<0.01 & 0.001. (B) Expression of radioresistance markers in radioresistant oral sublines. Western blotting for Cox-2, Bcl-2, Mcl-1 and Survivin proteins; β-Actin is used as loading control. Densitometry analysis of western blots from three separate experiments is shown below, bars represent mean ± s.d. * represent protein expression compared to parental cell line, while # represent protein expression compared between 50Gy-UPCI:SCC029B and 70Gy-UPCI:SCC029B sublines (p<0.05).
Figure 2.
Fractionated irradiation leads to change in cell morphology.
(A) Cell morphology analysis. Parental UPCI:SCC029B cells, radioresistant 50Gy-UPCI:SCC029B and 70Gy-UPCI:SCC029B cells. Scale bars: 100 µm. (B) Confocal images of filamentous-Actin stained with Alexa Fluor-488 phalloidin, cells were counter stained with DAPI. Scale bars: 20 µm. Arrow indicates filopodia as fine cell surface extensions. Analysis based on 50 cells per population with mean and standard deviation of three independent experiments, plotted at below right side (p<0.01, 0.001).
Figure 3.
Mean Raman spectra of parental and radioresistant sublines.
(A) Parental UPCI:SCC029B cell line (B) 50Gy-UPCI:SCC029B subline (C) 70Gy-UPCI:SCC029B subline.
Figure 4.
Difference Raman spectra of parental and radioresistant sublines.
(A) 50Gy-UPCI:SCC029B subline – parental UPCI:SCC029B cell line (B) 70Gy-UPCI:SCC029B subline – parental UPCI:SCC029B cell line (C) 50Gy-UPCI:SCC029B subline –70Gy-UPCI:SCC029B subline.
Figure 5.
PCA analysis for parental and radioresistant sublines.
(A) Loads of factor 1, 2 and 3 (B) 3-D scatter plot for parental UPCI:SCC029B cell line, radioresistant 50Gy-UPCI:SCC029B and 70Gy-UPCI:SCC029B sublines.