ABSTRACT
Objective To explore the effect of S100 calcium ion binding protein A6 (S100A6) on proliferation and migration of esophageal adenocarcinoma SK-GT-4 cells. Methods Lenti viruses were used to construct stable transfected cell lines (shNC and shS100A6). Real-time PCR was used to detect the mRNA expression of S100A6. The inverted microscope and MTT were used to detect cell proliferation. The Transwell assay was used to detect cell migration. Western blotting was used to detect the expression of S100A6, p-ERK, p-Akt and its downstream molecular involved in proliferation and migration. Using U0126 ( inhibitor of MER1/2) and LY294002 ( inhibitor of PI3K) to detect the effect of these two inhibitors on cell proliferation and migration and the expression of p-ERK, p-Akt and its downstream molecular involved in proliferation and migration in shS100A6 cells. Results Stable cell lines of knockdown S100A6 were constructed. Knockdown S100A6 promoted cell proliferation and migration. Western blotting result displayed that in shS100A6 cells, the levels of p-Akt and p-ERK increased, p21 decreased, cyclinDl increased, and the expression of β-catenin and vimentin, increased. U0126 and LY294002 inhibited the migration of shS100A6 cells. U0126 had no effect on the proliferation of shS100A6 cells, however LY294002 could inhibit the proliferation of shS100A6 cells. U0126 treatment on shS100A6 cells could decrease p-ERK and β-catenin expression. After shS100A6 cells treated with LY294002, p-Akt and β-catenin expression decreased, p21 expression increased and the expression of cyclinDl decreased. Conclusion Low expression of S100A6 promotes cell proliferation and migration, which may be mediated by activation of p-Akt regulating cell cycle progression to promote cell proliferation and by activation of p-Akt/p-ERK to regulate β-catenin to promote cell migration.
ABSTRACT
Objective To explore the effect of oridonin (ORI) on proliferation, apoptosis, cell cycle and migration of esophageal squamous carcinoma cell (ESCC) lines KYSE-150 and KYSE-450. Methods The effect of ORI on the proliferation and clony formation of esophageal cancer cells were detected by MTT and colony formation assays. Flow cytometry was performed to examine the impact of ORI on cell apoptosis and cell cycle. Transwell assay was applied to detect the role of ORI on cell migration. The effect of ORI on the expression of anti-apoptotic protein Bcl-2, cell cycle inhibitory protein p21Cip1/Waf1, epithelial-mesenchymal transition (EMT) related markers were examined by Western blotting. Results ORI had a significant inhibitory effect on the proliferation, migration and clone formation of KYSE-150 and KYSE-450 cells (P<0. 05) in a time and dose-dependent manner. With the increase of ORI concentration, apoptosis rate and the proportion of cells in G2/M phase increased significantly (P<0. 05), and the proportion of cells in G0/G1 phase decreased significantly (P < 0. 0 5). Bcl-2, vimentin and p-catenin were down-regulated and p21Cipl/Wafl, Ecadherin were up-regulated after treatment of ORI on ESCC cells for 48 hours. Conclusion ORI may inhibit ESCC cell proliferation and clony formation by inducing apoptosis and resting cells in G2/M phase, and suppress ESCC cell migration via inhibiting EMT process.
