The Anti-tumor Effect of Macrolides on the Extracellular Matrix Invasion and Cell Adhesion in Human Non-small Cell Lung Cancer Cells

ABSTRACT

PURPOSE: We investigated the effect of macrolides on the extracellular matrix (ECM) invasion and expression of cell adhesion proteins in non-small cell lung cancer cells. MATERIALS AND METHODS: Three kinds of macrolides, azithromycin, erythromycin and clarithromycin were treated in several human Non-Small Cell Lung Cancer (NSCLC) cell lines at doses from 0.1 to 1.5microg/ml. ECM invasion was measured by an in vitro chemo-invasion assay using matrigel-coated invasion chambers. RESULTS: Azithromycin inhibited the migration of NCI-H157 and NCI-H1299 cell lines. Erythromycin inhibited the migration of NCI-H157 and NCI-H2066, and clarithromycin had inhibitory effects on the migration of NCI-H358 and NCI-H2009. These results indicate that macrolides inhibit the ECM invasion of the tested NSCLC cells cell line-specifically. Western blot analyses for cell adhesion proteins (CAPs) showed that these were up-regulated by treatment with macrolides in some of the NSCLC cell lines. Based on the results of chemo-invasion assay, we selected each 2-cell line group, one was significantly suppressed by AM (NCI-H157 and NCI-H1299) and the other was less effectively suppressed by EM (NCI-H358 and NCI-H460). The expression levels of cell adhesion proteins increased in a dose-dependent manner in two cell lines in which their ECM invasion was inhibited by AM, however, the other two cell lines showed unchanged expression levels even at higher doses. Furthermore, the expressions of E1AF and matrix metalloproteinases (MMPs) activity were examined by RT-PCR and gelatin zymography, respectively with the selected 4 cell lines. E1AF expression level was found to decrease from the basal level in NCI-H157 and the activity of MMP-9 was also shown to decrease in NCI-H157 and NCI-H1299. Cell adhesion to fibronectin was a little reduced in NCI-H157 from its basal level. CONCLUSION: Consequently, the present study suggests that macrolides inhibit the ECM invasion of NSCLC cells by inducing the altered expression of cell adhesion proteins