The Influences Of TGF-β1Upon The Human Adenocarcinoma Cell Of Lung A549and Peripheral Blood Mononuclear Cells

【Objective】 To explore the change of cell morphology and cellular immunity influences of TGF-β1upon the human lung adenocarcinoma cell A549and peripheral blood mononuclear cells,.[Method] Culture of lung adenocarcinoma cell A549line and stably propagated in vitro. then adding different concentration (2ng/ml,5ng/ml,10ng/ml,20ng/ml) of TGF-β1and the mononuclear cells isolated from peripheral venous blood of NSCLC patients, to co-culture. Use inverted phase contrast microscope to observe the cell morphology and flow cytometry to detect the concentration of IL-2,IL-4,IL-6, IL-10,TNF and INF-gamma, and count CD3+,CD4+,CD8+,CD3+CD25+,CD4+CD25+cell proportion in a sequence time point of3day,5day,8day.[Results]1. The TGF-β1group A549cells' morphology change the cobblestones tightly junction-look growth pattern to elongated shuttle shape connected loosely-look growth pattern compared with non-TGF-β1group. And A549cells' morphology deviation increased with the raised concentration and prolonged treatment of TGF-β1.2. The TGF-β1group PBMCs' morphology change the suspended cluster-look growth pattern to diffuse isolated-look growth pattern and the diffuse floating cells appeared early compared with non-TGF-β1group. With the increase of concentration and prolonged treatment of TGF-β1, the effect increase.3. The cytokines' concentration of A549cells supernatant's in the TGF-β1(2ng/ml, 5ng/ml,10ng/ml,20ng/ml) groups at a sequence time point of3day,5day,8day.TGF-β1group impact on the Thl cytokines' concentration of A549cells: IL-2,TNF,IFN-γ is lower than non-TGF-β1group(P<0.05).For the Th2cytokines' concenttration of A549cells: IL-4,IL-10is higher than non-TGF-β1group(P<0.05).In cenain range,with the increase concentration and prolonged treatment of TGF-β1.the effect raised.IL-6shows no significant different between TGF-β1group and non-TGF-β1group at sequence time point(P>0.05).4. The cellular immunity of PBMCs in the TGF-β1(2ng/ml,5ng/ml,10ng/ml,20ng/ml) groups at a sequence time point of3day,5day,8day.In TGF-β1groups the proportion of CD3+,CD4+,CD8+,CD3+CD25+cells decreased compared with non-TGF-β1group(P<0.05),and the proportion of CD4+CD25+cells raised significantly(P<0.01). In certain range, with the increase concentration and prolonged treatment of TGF-β1,the effect raised.[Conclusion]1. During the co-culture adenocarcinoma cells of lung with PBMCs,TGF-β1can induced human lung cancer cells show epithelial-to-mesenchymal transition. Enhanced its ability of migracion and infiltration.Simultaneously,TGF-β1can depressing the growth and proliferation of PBMCs,inhabiting T-cell activation, and accelerating the PBMCs apoptosis.2. TGF-β1can inhabit adenocarcinoma cells of lung Th1related-cytokines, enhance Th2related-cytokines,cause the disorder of Th1/Th2,resulting in the Th1cellular dominate immunity decline.3. TGF-β1may affect related-cytokines secretion to inhabit the activation of T-cells, Cause the dysfunction of immune surveillance and cytotoxic T-lymphoCyte.Result in the inhibition of cellular immunity.