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Objective_To study the effect of Y-27632 on invasion and motility of SGC-7901 gastric carcinoma cells, and to find whether Y-27632 excerts the effect by attenuating SRF expression.Methods_SGC-7901 gastric carcinoma cells were divided into 3 groups:1)blank control group;2)Y-27632 group;3)siRNA-SRF-1107 group. Transfected siRNA-SRF or incubated by Y-27632 48 h.The effect of Y-27632 on proliferation suppressions of SGC-7901 gastric carcinoma cells was detected by CCK-8 assay.Cell invasion was examined by Transwell and wound healing test.The expression of SRF, ROCK1, E-cadherin, β-catenin, F-actin, MRTF-A and Cyclin D1 were detected by Western blot.Results_Y-27632 inhibited invasion (P<0.05)but had no effect on proliferation of SGC-7901 gastric carcinoma cells.Y-27632 reduced ROCK1, MRTF-A, F-actin, SRF protein expressions by 37.0%, 44.3%, 62.7%and 62.7%respectively, and E-cadherin protein expression was up-regulated by 2.64 folds(P<0.05).Conclusions_The inhibition of ROCK and up-regulation of E-cadherin by Y-27632 can inhibit the invasion and migration of SGC-7901 gastric carcinoma cells that is explained at least, in part, by attenuating SRF expression.
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Objective:To construct pEGFP-C1-Smad4 expression vector and to observe the influence of Smad4 over-expression on the proliferation of human gastric cancer SGC7901 cells and its relationship with nuclear factor kappa B (NF-?B). Methods:Recombinant expression vector pEGFP-C1-Smad4 was constructed and was used to transfect human gastric cancer SGC7901 cells. EGFP expression in transfected cells was detected by fluoroscopy. Smad4 and NF-?B expression in transfectant was examined by Western blotting. Effect of Smad4 over-expression on activation of NF-?B and proliferation of transfected SGC7901 cells were examined by electrophoretic mobility shift assay (EMSA) and MTT assay,respectively. Results:Expression of EGFP in transfected SGC7901 cells was observed under fluorescence microscope. Smad4 was over-expressed in transfected SGC7901 cells,accompanied by down-regulation of NF-?B p65 expression in the tranfectants. EMSA and MTT demonstrated that Smad4 over-expression significantly inhibited the activation of NF-?B and the proliferation of SGC7901 cells (P
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The experiment was carried out on cultured human stomach glandular carcinoma (SGC-7901 cell line). We previously reported the cytochemical changes of (Na~+-K~+)-ATPase activity and the changes of microvilli. We here studied the relationship among the cAMP, cAMP-PDEase, Mg~(++)-ATPase activity, the cell growth and differentiation. Cells were exposed to Immol/L cAMP together with 1 mmol/L theophylline. We showed cAMP specific fluorescence by immunocytochemical method, Mg~(++)-ATPase and cAMP-PDEase activity by cytochemical methods. We found that cAMP with theophylline not only inhibited the growth of these SGC-7901 cells as shown by our previous result, but also decreased the activity of Mg~(++)-ATPase and 3', 5'-cyclic monophosphate phosphodiesterase. The result indicates that exogenous cAMP can inhibit 3', 5'-cAMP-PDEase and Mg~(++)-ATPase activity and increase cAMP level in cell, and cAMP regulated the growth and differentiation of SGC-7901 cell line