ABSTRACT
Objective To investigate the microenvironment alteration in epithehiae-mesenchymce transition(EMT)metastasis in gastric carcinoma induced by transforming growth factor β(TGF-β). Methods The gastric carcinoma cell line NCI-N87 was treated with TGF-βto induce cells to undergone EMT,real-time quantitative PCR(RT-qPCR)and immunofluorescence staining were used to examine expression of EMT markers and wound-healing assays,and transwell migration and invasion assays were performed to determine the potential of cell migration and invasion. Further,alteration of cytokines in tumor microenvironment was detected using real-time quantitative PCR and ELISA. Moreover,the activity of its downstream signaling molecules was detected by Western blot. Results TGF-β induced gastric carcinoma cells NCI-N87 to undergone EMT as well as promote cell migration and invasion. Further,TGF-βinduced upregulation of epidermal growth foctor(EGF)and vascular endothelial growth facfor(VEGF)expression,as well as downregulation of Dickkopf-1(DKK1)and secreted frizzled receptor proein1(SFRP1),which activated PI3K/AKT and Wnt/β-catenin sequentially. Conclusion TGF-β promotes EMT by inducing microenvironment alteration in gastric carcinoma cell NCI-N87.
ABSTRACT
Objective: To investigate the microenvironment alteration in epithehiae-mesenchymce transitionEMT metastasis in gastric carcinoma induced by transforming growth factor βTGF-β.
ABSTRACT
The anti-proliferative efficacy of t,t-conjugated linoleic acid (t,t-CLA), c9,t11-CLA, and t10,c12-CLA was compared in several human cancer cell lines. Gastric NCI-N87, liver Hep3B, pancreas Capan-2, and lung NCI-H522 cancer cells were incubated with 50 microM CLA isomers over a period of 6 days. The t,t-CLA inhibited the growth of all cancer cell lines to different extents, but c9,t11-CLA and t10,c12-CLA inhibited or stimulated the growth of the cancer cell lines. NCI-N87 cells were the most sensitive to growth inhibition and apoptosis from all CLA isomers tested. In NCI-N87 cells, CLA isomers reduced the release of arachidonic acid (AA) via the inhibition of cytosolic phospholipase A2 (cPLA2) activity, consequently reducing the production of PGE2 through the inhibition of cyclooxygenase-2 (COX-2). The efficacies of CLA isomers were in the following order (from most to least effective): t,t-CLA, t10,c12-CLA and c9,t11-CLA. Overall, these results imply that the anti-proliferative efficacy of t,t-CLA on cancer cells, especially NCI-N87 cells, was greater than other CLA isomers due to its induction of apoptosis through the inhibition of cPLA2 and COX-2 activities.
Subject(s)
Humans , Apoptosis , Arachidonic Acid , Cell Line , Cyclooxygenase 2 , Cytosol , Dinoprostone , Linoleic Acid , Liver , Lung , Pancreas , Phospholipases A2ABSTRACT
BACKGROUND/AIMS: The unavailability of human gastric cell lines representative of the normal gastric epithelial function such as polarized monolayer restricts the application of cell culture system in approaching the field of Helicobacter pylori (H. pylori) infected gastric mucosa models. The present investigation aimed at assessing the usefulness of NCI-N87 cell line as an adequate cellular model to study the pathophysiology of human H. pylori infection. METHODS: For the identification of epithelial phenotypes at low magnification, cells were observed on a phase-contrast microscope and confocal microscope. Transepithelial resistance (TER) was measured on NCI-N87 cells seeded on Transwell(R) to identify monolayer polarity two or three times a week after confluency. The IL-8 level was determined by ELISA at 24 hours after the administration of HP60190 and IL-1alpha on NCI-N87 cells. IL-8 level was compared in both upper and lower well with the control. RESULTS: A monolayer phenotype was observed in NCI-N87 cell lines by using confocal microscope. TER was measured as 400-500 (omega x cm2) at two or three weeks after cell culture. In NCI-N87 cell lines, IL-8 level was significantly increased after 24 hour compared to control, and was prominent in the lower well. CONCLUSIONS: These results suggest that NCI-N87 cell line may be useful in H. pylori infected gastric mucosa model.