ABSTRACT
<p><b>BACKGROUND</b>Tumor cells can reduce the number of dendritic cells (DCs) in the tumor environment and cause DC dysfunction through autocrine or paracrine pathways. We sought to measure cyclooxygenase-2 (COX-2) expression in bombesin-inhibited DCs treated with theanine in vitro and to explore the protection and activation effects of theanine on DCs.</p><p><b>METHODS</b>Enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting were used to analyze the effects of theanine on COX-2 expression and interleukin (IL)-12/IL-10 secretion of bombesin-treated DCs.</p><p><b>RESULTS</b>DCs acquired an impaired phenotype as a result of bombesin treatment. Theanine increased the expression of mature DC surface molecules. The number of cell apoptosis with the treatment of bombesin and theanine significantly decreased, accounting for 15.9%, compared with 26.1% of cell apoptosis with bombesin. COX-2 expression in bombesin-treated DCs was inhibited by theanine in a dose-dependent manner. Theanine promoted DC secretion of IL-12. IL-12 levels reached (137.4 ± 4.9) pg/ml with theanine at 200 µmol/L. However, theanine inhibited the secretion of IL-10 in a dose-dependent manner. IL-10 levels were only (58.4 ± 6.9) pg/ml with theanine at 200 µmol/L.</p><p><b>CONCLUSION</b>Theanine inhibits the transcription and translation of COX-2 and regulates the balance of IL-10/IL-12 secretion in bombesin-inhibited DCs, leading to the recovery of a state of activation in DCs.</p>
Subject(s)
Humans , Bombesin , Pharmacology , Cells, Cultured , Cyclooxygenase 2 , Metabolism , Dendritic Cells , Metabolism , Enzyme-Linked Immunosorbent Assay , Glutamates , Pharmacology , Interleukin-10 , Metabolism , Interleukin-12 , MetabolismABSTRACT
<p><b>BACKGROUND</b>Evidence shows that ezrin plays an important role in the development of some human malignancies. But the mechanism by which ezrin may affect tumor cell invasion and metastasis remains unclear.</p><p><b>METHODS</b>In this study, the expression of ezrin was verified in osteosarcoma (OS) cells and tissues by comparison with normal bone cells and tissues using Western blotting. OS-MG63 were transfected with pcDNA3.1-ezrin or pGenesil-1/shRNA-ezrin and the stably transfected cells were selected with G418 to yield the ezrin cell line. The OS-MG63 tumor cells were delivered by tail vein to female BALB/c to develop pulmonary metastasis model in vivo. Ezrin was identified as a direct target of miR-183 via a luciferase reporter carrying the 3'-untranslated region of ezrin. Migration assays and invasion assays were done with the transwells. Signaling pathway was studied by Western blotting and/or inhibitor.</p><p><b>RESULTS</b>Ectopic overexpression of ezrin in OS cell line MG63 promoted tumor cell invasion and migration. Consistent with this, knockdown of ezrin inhibited tumor cell invasion and migration. Similar results were obtained in the experimental metastasis model in vivo. We identified ezrin as a direct target of miR-183. What is more, ectopic expression of ezrin could induce the expression of N-cadherin and enhance the activity of extracellular signal-regulated kinase (ERK) signaling.</p><p><b>CONCLUSION</b>Collectively, these results suggest that ezrin as a direct target of miR-183 promotes the aggressiveness of OS via increased N-cadherin and activating ERK signaling.</p>
Subject(s)
Animals , Female , Humans , Mice , Bone Neoplasms , Metabolism , Pathology , Cadherins , Genetics , Metabolism , Cell Line, Tumor , Cell Movement , Genetics , Cytoskeletal Proteins , Genetics , Metabolism , In Vitro Techniques , Neoplasm Invasiveness , Genetics , Osteosarcoma , Metabolism , PathologyABSTRACT
Objective To reveal the impact of anti-lung cancer A549 by theanine, which may regulate T lymphocytes through dendritic cells (DCs), and to observe the immune protection of theanine on SCID mice transplanted tumors. Methods SCID mice were reconstructed after immunization, then A549 cells were inoculated. The immune protection and immune treatment of theanine stimulated DCs were observed. Results In the theanine-stimulated DC group, as well as DC group, the time needed to form tumor were significantly prolonged (F = 29. 5, P ＜ 0. 01). And the transplanted tumors were smallest (F =69. 51, P ＜0. 01) and lightest (F = 190. 9, P ＜0. 01) in the theanine-stimulated DC group. 50 days after tumor-bearing, the transplanted tumors in the theanine-stimulated DC group were smaller than in the control group, and the surface were smoother with no adhesions. Under light microscope, a large thin slice necrosis was showed in the theanine-stimulated DC group, and the VEGF expression was also reduced (F = 31.4, P ＜ 0. 01). Conclusions The anti-tumor immune protection and treatment mechanism of theanine-stimulated DCs were possibly through regulation of T cells, as well as the VEGF expression reducing,thereby inhibited tumor blood vessel formation.
ABSTRACT
Objective To observe the effect of theanine in vitro and in vivo, including suppression of lung tumor growth, tumor an-giogenesis and promotion of apoptosis. Methods The inhibitory effects of theanie on lung cancer A549 cells were analyzed by MTT assays. The cell cycle and the apeptotic percentage of A549 cells were detected by FCM. The angiogenesis effect of theanine was observed with CAM model. The inhibitory effects of theanine were observed with lung carcinoma nude mice model, and the immunohistochemic technique was used to investigate the expression of CD34 and VEGF (vascular endothelial growth factor). Results Treatment of A549 cells with VEGF re-sulted in significant dose-dependent and time-dependent inhibition. FCM detection indicated that administration of EGCG resulted in an in-crease in cells in the S phase of the cell cycle and a typical apoptosis peak before the GI phase with an apoptosis rate of 15.9%. There was a significant difference in tumor volume and weight in theanine group compared with control group after two-week treatment, and the tumor in-hibition rate was 43.6%. There was no significant difference in expression of VEGF in tumor tissue according to tumor MVD marked by CD34 between the theanie group and control group. Thus, theanine could obviously inhibit tumor angiogenesis. Conclusion VEGF can ar-rest lung tumor growth via the promotion of tumor cell apoptosis and the inhibition of tumor angiogenesis.
ABSTRACT
Objective To evaluate the role of IL-2 dendritic cell in chronic hepatitis B,peri-dendritic cells were cultured,cytokins CD4O,CD86,CD80,CD83 and IL-2 were measured by ELLISA.Methods 20 chronic hepatitis B patients were selected,each one donated 15ml blood.dentritic cells were cultured,and IL-4 1000u/ml rhGM-CSF 1000u/ml was given each well,while IL-2 100u/ml was given each control well.CD40,CD86,CD80 and CD83 were measured by FLSC at 9th day.IL-2 was detected by ELISA at 12th day.Results The level of CD40,CD86,CD80 and CD83 in increased when using IL-2,and dentritic cell secreted IL-2 in chronic hepatitis.Conclusion The results suggest that IL-2 can promote the phenotypic and functional maturation of PC.