## 3 Results  
PrestoBlue(tm) cell viability test was conducted to determine the optimal concentration of nifedipine to be used for cell treatment that did not significantly affect cell viability. PrestoBlue(tm) data showed that nifedipine, regardless of treatment concentration, had no effect on cell viability compared to that of untreated cells (Figure 1). Considering the percentage of viability cells, the concentration of 1000 ng/ul was assumed as not lethal but sufficient to induce a significant response. Based on this test, the optimal concentration used for the treatment was 1000 ng/mL. The gene expression profile of 57 genes belonging to the \"Extracellular Matrix and Adhesion Molecules\" pathway was analyzed using real-time PCR. Table 1 reports the list of genes and their fold change after treatment with nifedipine. Bold fonts indicate significant variation of gene expression level: fold change \(\geq\) 2 and \(p\)-value \(\leq\) 0.05 for upregulated genes, and fold change \(\leq\) 0.5 and \(p\)-value \(\leq\) 0.05 for significantly downregulated genes. Figure 1: Nifedipine effect on cell viability. \begin{table}
\begin{tabular}{c c c c} \hline \hline
**Gene** & **Fold Change** & **SD** & **Gene Function** \\ \hline CD44 & 0.64 & 0.08 & Cell-Cell Adhesion \\ \hline CDH1 & **5.62** & 2.06 & Cell-Cell Adhesion \\ \hline COL1A2 & 0.77 & 0.12 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL2A1 & 0.57 & 0.15 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL3A1 & 0.66 & 0.05 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL4A1 & 0.85 & 0.14 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL5A1 & 0.66 & 0.08 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL6A1 & 0.90 & 0.17 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL7A1 & 0.55 & 0.19 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL8A1 & 0.92 & 0.09 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL9A1 & 1.10 & 0.02 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL10A1 & 1.54 & 0.06 & Collagens and Extracellular Matrix Structural constituent \\ \hline COL11A1 & 0.96 & 0.11 & Collagens and Extracellular Matrix Structural constituent \\ \hline CCTNA1 & 0.69 & 0.02 & Cell Adhesion Molecule \\ \hline CTNB & 0.67 & 0.07 & Cell Adhesion Molecule \\ \hline CTNND2 & 0.96 & 0.30 & Cell Adhesion Molecule \\ \hline FN1 & 0.54 & 0.09 & Cell Adhesion Molecule \\ \hline HAS1 & 0.68 & 0.09 & Transmembrane Receptor \\ \hline ILF3 & 0.62 & 0.07 & Transmembrane Receptor \\ \hline ITGA1 & 0.95 & 0.10 & Transmembrane Receptor \\ \hline ITGA2 & 1.19 & 0.18 & Transmembrane Receptor \\ \hline ITGA3 & 0.88 & 0.15 & Transmembrane Receptor \\ \hline ITGA4 & 0.61 & 0.10 & Transmembrane Receptor \\ \hline ITGA5 & 0.70 & 0.03 & Transmembrane Receptor \\ \hline ITGA6 & **0.41** & 0.04 & Transmembrane Receptor \\ \hline ITGA7 & 1.48 & 0.31 & Transmembrane Receptor \\ \hline ITGA8 & 0.59 & 0.06 & Transmembrane Receptor \\ \hline ITGB1 & 0.87 & 0.09 & Transmembrane Receptor \\ \hline ITGB2 & 0.73 & 0.07 & Transmembrane Receptor \\ \hline ITGB4 & **0.23** & 0.03 & Transmembrane Receptor \\ \hline ITGB5 & 0.75 & 0.06 & Transmembrane Receptor \\ \hline LAMA1 & **0.45** & 0.04 & basement Membrane Constituent \\ \hline LAMA2 & 1.06 & 0.19 & basement Membrane Constituent \\ \hline LAMA3 & 0.64 & 0.08 & basement Membrane Constituent \\ \hline LAMB1 & **0.39** & 0.02 & basement Membrane Constituent \\ \hline LAMB2 & 0.75 & 0.10 & basement Membrane Constituent \\ \hline LAMB3 & 0.88 & 0.11 & basement Membrane Constituent \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Genes belonging to the “Extracellular Matrix and Adhesion Molecules” pathway analyzed using real-time Polymerase Chain Reaction. Fold changes represent the expression levels of genes after 24 h treatment with nifedipine, as compared with untreated cells (\(n=3\)). SD = standard deviation.