Influence Of Interactions Between Lung Cancer Cells,Inflammatory Cell Or Fetal Lung Fibroblast On Matrix Metalloproteinase-1, -2, -9 Expression And Inhibiting Effects Of Retinoic Acid On Lung Tissue Destruction

Intereractions between tumor cells and host components,such as immune cells, fibroblasts, and endothelial cells, contribute to the progression of cancer from its initial growth at a primary site in the body to its metastasis to distant organs. Matrix metalloproteinase(MMPs) are a family of zinc-dependent endopeptidases that have been shown to contribute to tumor progression and metastasis. Both tumor and host-derived MMPs contribute to multiple stages of tumor progression through their ability to degrade the basement membrane and components of the extracellularmatrix. To observe the effects of the co-culture of H460 with mononuclear cells and HFL in three-dimensional (3D) collagen gels on the expressions of matrix metallop roteinase-1,-2,-9 (MMP-1,-2,-9) in the first and second study.Emphysema is a condition of the lung characterized by abnormal, permanent enlargement of airspaces distal to the termal bronchiole, accompanied by destruction of their walls and without obvious fibrosis.Proteinase-Antiproteinase imbalance is a primary inducement which leads to elastin and alveolar structure destruction.Recently many evidences suggest that there is daedal extracelluar matrix remodeling in the development of emphysema.Increased extracelluar matrix collagen deposition and abnormal collagen remodeling are a novel view point in the pathogenesis of emphysema. the embalance of MMPs/TIMPs secretion and the lose control of their activation play an important role in the process. So far, there is no available drug therapy to control COPD. Recently studies find that retinoic acid (RA) can decrease the expression of collagenase, promote impaired epithelium reparation and increase elastin synthesis, which suggest that RA is beneficial to control COPD. To investigate the effects of neutrophil elastase (NE) and all-trans-retinoic acid in lung fibroblast mediated collagen degradation in three dimensional culture.The experiment included three parts: 1. Influence of interactions between H460 and mononuclear cells on matrix metalloproteinase-1,-2,-9 expressionAIM:To observe the effects of the co-culture of H460 with mononuclear cells in three-dimensional (3D) collagen gels on the expressions of matrix metalloproteinase-1,-2,-9(MMP-1,-2,-9).METHODS:Cells cultured in 3D collagen gels were divided into the control group(CG), the H460 group (HG), the mononuclear cell group (MG) and the co-culture group of H460 with mononuclear cells (HMG), respectively. The growth of cells was observed under microscope. The expressions of MMP-2,-9 were detected by gelatin zymography at 24,72,120 h, and MMP-1 were detected by Western blotting.RESULTS:Cells in collagen gels grew well, and MMP-1,-2,-9 were not detected in MG, but could be obviously seen in HG and HMG. The levels of MMP-1,-2,-9 in HMG were significantly higher than those in HG and CG.CONCLUSION:The results demonstrate that MMPs could be induced by the interaction between cancer cells with inflammatory cells, and MMPs may promote the invasion and metastasis of cancer cells by degrading collagen and the ECM of cancer tissue.2. Influence of interactions between lung cancer cells%fetal lung fibroblast and inflammatory cell on matrix metalloproteinase-1,-2,-9 expressionAIM:To observe the effects of the co-culture of H460 with mononuclear cells and HFL in three-dimensional (3D) collagen gels on the expressions of matrix metallop roteinase-1,-2,-9 (MMP-1,-2,-9).METHODS:Cells cultured in 3D collagen gels were divided into the control group(CG),the H460 group (HG),the mononuclear cell group (MG),the fetal lung fibroblast group (FG),the co-culture group of H460 with mononuclear cells (HMG),the co-culture group of H460 with HFL (HFG), and the co-culture group of H460 with mononuclear cells and HFL(HMFG) respectively. The growth of cells was observed under microscope. The expressions of MMP-2,-9 were detected by gelatin zymography, and the expressions of MMP-1 were detected by Western blotting.RESULTS:Cells in collagen gels grew well, and MMP-1,-2,-9 were not detected in CG and MG, but could be obviously seen in HG,FG,HMG,HFG and HMFG, and the levels of MMP-1,-2,-9 in HMFG were significantly higher than those in the other 4 groups.3. Inhibiting effects of retinoic acid on lung tissue destruction in three imensional culture of human lung fibroblasts in type I collagen gelsAIM:To investigate the effects of neutrophil elastase (NE) and all-trans-retinoic acid in lung fibroblast mediated collagen degradation in three dimensional culture.METHODS:Human lung fibroblasts were cultured in three dimensional collagen gels. TNF-αand IL-βwere added to induce MMPs, and RA or NEwas added to the culture media at same time.The expressions of MMP-2,-9 were detected by gelatin zymography, and MMP-1,-3 and TIMP-1,-2 were detected by Western blotting. Collagen content and gel areas were determined on day 5.RESULTS:TNF-αand IL-βinduced fibroblast productions of MMP-1,3, 9 in latent forms. NE cleaved the inhibitors of MMP and activated MMPs into active forms, resulting in generously collagen degradation.The conclusions were as follows:1. The results demonstrate that MMPs could be induced by the interaction between cancer cells with inflammatory cells, and MMPs may promote the invasion and metastasis of cancer cells by degrading collagen and the ECM of cancer tissue.2. The results demonstrate that more MMPs were secreted in HMFG than in the other 5 groups, and mononuclear cells and HFL in carcinoma tissue may promote the invasion and metastasis of cancer cells; MMPs may promote the invasion and metastasis of cancer cells by degrading collagen and the ECM of cancer tissue.3. The results demonstrate that the synergistic interaction between neutrophil elastase and MMPs may be involved in the lung tissue destruction in emphysema and chronic obstructive pulmonary disease, however,ATRA may against the effect of NE in the lung tissue destruction in emphysema and chronic obstructive pulmonary disease.