A Preliminary Study Of The Nitric Oxide In Promoting Lung Cancer Invasion And Metastasis

Objective:The aim of this study is to elucidate the relationship between the expressions of iNOS, cytoskeleton protein caldesmon, OPN, and clinical parameters especially the metastasis of lung cancer.Methods:(1) The change of the concentration of intracellular calcium in cell A549after treated with SIN-1treatment or iNOS transfection was analyzed by FCM.(2) Western blotting was applied to detect the expression of caldesmon in cell A549after treated with SIN-1treatment or iNOS transfection.(3)The distribution and quality change of cell A549after treated with SIN-1treatment or iNOS transfection was identified by immunofluorescence stain.(4) The migration ability of cell A549after treated with SIN-1treatment or iNOS transfection was detected by wound healing assay.(5) The distribution and expression of iNOS, caldemon, OPN in lung cancer tissue and intracranial metastatic tumor were detected by Immunohistochemistry.(6)The ratio of EPCAM+/OPN in blood serum of NSCLC patients with and without bone metastasis was analysed by FCM.(7)The concentration of OPN in blood serum of NSCLC patients with and without bone metastasis was measured by ELISA.Results:(1) FCM showed that the concentration of intracellular calcium in cell A549after treated with SIN-1treatment or iNOS transfection was up-regulated.(2) Western blotting showed that the expression of caldesmon was up-regulated in A549cells after they were treated with SIN-1treatment or iNOS transfection.(3) Immunofluorescence stain showed that caldesmon were localized at the edge of A549cells and formed punctate F-actin-enriched structures. The iNOS inhibitor1400W can block caldesmon polymerization.(4) The wound healing assay showed that SIN-1treatment or transfection with iNOS obviously increased the rate of wound closure of A549in a time-dependent manner.(5) OPN was expressed in tumor cells; iNOS was expressed mainly in stromal cells near the tumor tissues Caldesmon was expressed in tumor tissues and the expression of iNOS, caldemon, OPN was up-regulated.(6) FCM showed that the ratio of EPCAM+/OPN in blood serum of NSCLC patients with bone metastasis was higher than the without bone metastasis ones.(7) ELISA showed that the concentration of OPN in serum of NSCLC patients with bone metastasis was higher than the without bone metastasis-ones.Conclusion:These results suggested that iNOS, caldesmon and OPN may work as biomarkers for metastasis of lung cancer. This article include8figures,4tables,102references.