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

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

Effect of Uteroglobin on cPLA2, COX-2 Expression and ERK Activation in NSCLC Cells / 결핵

BACKGROUND: Uteroglobin is a protein produced by the normal bronchial epithelium and its expression level is lower in non-small cell lung cancer tissues and cell lines. It mainly functions as an anti-inflammatory, and when it is overexpressed in cancer cells, the neoplastic phenotype is antagonized. cPLA2 and COX-2, which are also associated with inflammation, were reported to be related to cancer. The relationship between cPLA2, COX-2 and uteroglobin is unclear. The relationship between uteroglobin and ERK, which is related to cell growth, is also not unclear. This study investigated the changes in the cPLA2 and COX-2 expression levels and the ERK activities after the overexpression of uteroglobin in non-small cell lung cancer cell lines. METHODS: The A549 and NCI-H460 cell lines were infected by adenovirus-null and adenovirus- uteroglobin. The cChange in the cPLA2, COX-2 expression level and ERK activity after uteroglobin overexpression was measured by Western blot. The change in MMP activity was measured by zymography. RESULTS: Western blot revealed decreased expression levels of cPLA2, and COX-2, and increased pERK levels in nonsmall cell lung cancer cells after uteroglobin overexpression. Zymography revealed no changes in the MMP-2 activity and lower MMP-9 activity. U0126, which is a specific inhibitor of ERK-activating kinase MEK-1/-2, prevented the decrease in the MMP-9 activity CONCLUSIONS: A decrease in cPLA2 expression, COX-2 expression, MMP-9 activity and a increase in ERK activity may be related to the anticancer effects of uteroglobin in nonsmall cell lung cancer cells.

The Expression of Clara Cell Secretory Protein in BAL Fluid of Patients with Idiopathic Interstitial Pneumonia / 결핵

BACKGROUND: Idiopathic interstitial pneumonia is characterized by chronic inflammation and pulmonary fibrosis. The clara cell 10 kD protein (CC10, also designated CC16) is synthesized by the bronchial epithelium and has been suggested to have a potent anti-inflammatory effect. Therefore, CC-10 might be a candidate for controlling the inflammatory events in patients with idiopathic interstitial pneumonia. The aim of this study was to determine if the degrees of pulmonary fibrosis in idiopathic interstitial pneumonia is associated with CC-10 in the BAL fluid. METHODS: The BAL fluid was collected from 29 patients and 10 controls. Densitometric analysis of the western blot assay for the CC-10 was subsequently performed. The RI (relative intensity) of each band was compared according to the diagnosis, the radiological degrees of pulmonary fibrosis and the relative proportion of inflammatory cells in the BAL fluid. RESULTS: There were no differences in the CC-10 expression levels in the BAL fluid between the patients (RI 77.5+/-75.8%) and the controls (70.7+/-39.8%) (p>0.05). In addition, the degrees of pulmonary fibrosis and airway inflammation in patients with usual interstitial pneumonia were not associated with CC-10 expression in the BAL fluid (p>0.05). CONCLUSION: This study suggests that CC-10 expression is not associated with the degrees of pulmonary fibrosis in patients with usual interstitial pneumonia.