Effects Of Interactions Between Interstitial Cells And Lung Cancer Cells On The Process Of Tumor's Invasion And Metastasis

Objrctive1. To explore the interactions among mononuclear cells,H460 and fetal lung fibroblast and the influence on the expression of IL-1βand TNF-αin three-dimensional culture model.2. To evaluate the influence of IL-1βon the expression of MMP-2 of HFL cells in three-dimensional culture collagen gels.3. To investigate the influence of different doses dexamethasone(DEX) of H460 cell growth stimulated by TNF-α.Methods1. Cells cultured in 3D collagen gels were divided into co-culture groups of mononuclear and H460 cells, mononuclear and fetal lung fibroblast, H460 and fetal lung fibroblast, mononuclear cells and H460 cells and fetal lung fibroblast,respectively.The growth of cells was observed under microscope.The expressions of IL-1βand TNF-αwere detected by ELISA at 24,72,96 h after culture.2. HFL cells were cultured in three-dimensional culture collagen gels. HFL ells were randomly divided into control and experimental groups. Cells of the control group were cultured in DMEM media without IL-1β.Cells of experimental groups were cultured in DMEM media with different concentrations of IL-1β(10,20 ng/ml),respectively. The phase contrast microscope was used to observe the morphological of the cells. After 24,72 hours, harvested the supernatant,MMP-2 were detected by gelatin zymography. The proliferation of HFL cell stimulated by IL-1βwas examined by tetrazolium salt (MTT) assay.3. The H460 were cultured and strimulated by TNF-αin vitro. Human non-small lung cancer of H460 cell were randomly divided into control and experimental groups.Cells of the control group were cultured in DMEM media without dexamethasone.Cells of experimental groups were cultured in DMEM media with different concentrations of dexamethasone (25,50,100,200μg/ml),respectively.The inhibitions of DEX of H460 cell stimulated by TNF-αwere examined by tetrazolium salt(MTT) assay. The phase contrast microscope was used to observe the morphological changes of the cellsResults1. Cells cultured in three-dimensional culture collagen gels were well. the levels of IL-1βand TNF-αwere enhanced in co-culture groups of mononuclear and H460 cells.The levels of IL-1βand TNF-αwere also enhanced slightly in co-cultured mononuclear cells and fetal lung.Co-cultures of fetal lung fibroblast with H460 cells resultured little IL-1βand TNF-α, When co-culture mononuclear cells and H460 cells and fetal lung fibroblast,the levels of IL-1βand TNF-αwere significantly higher than the other groups.2. Observation of the cells growing in three-dimensional co-culture:In three-dimensional co-culture, HFL cells were growing in gel-culture, and it didn't proliferate, and all of the cells were growing in singles;The expression of MMP-2 is different in different IL-1βconcentration, and the stimulus was enhanced with the increasing of IL-1βdoses;IL-1βcan enhance HFL proliferation in a concentration-dependent Mannes. The different concentrations of IL-1βhad different effects on pr oliferation of HFL (P<0.05), the proliferation ability was dose-time dependent.3. TNF-αcan enchance H460 proliferation in a concentration-dependent Manne.DEX(25～200μg/ml) inhibited of H460 growth in cultures with TNF-α(P<0.05). The different concentrations of DEX had different effects on proliferation of H460 (P<0.05), the proliferation ability was dose-time dependent.conclusions1. More IL-1βand TNF-αsecreted in co-culture mononuclear cells and H460 cells and fetal lung fibroblast.2. IL-1βmay enhance the invasion and metastasis of lung cancer though up-regulating the secretion and of MMP-2, and enhance HFL proliferation.3. DEX can inhibit H460 growth stimulated by TNF-αwhich may suggest a new approach to prevention and treatment of cancer.