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.
ABSTRACT
Plumbagin (Plumbago zeylanica L.) has a wide spectrum of anticancer activity with a relatively lower toxicity. The molecular mechanisms of proliferation inhibition and apoptosis induction by plumbagin on esophageal squamous carcinoma cell lines may be important for the structure modification and clinical application of plumbagin. After treatment of KYSE-30, KYSE-70 and KYSE-140 cells with 0-20 μmol·L-1 of plumbagin for 24, 48, 72 h, CCK8 was used to examine the proliferation, Annexin V and PI immunofluorescence staining for apoptosis, real-time PCR and Western blot for FoxM1 mRNA and protein expression, dual-luciferase reporter gene assay for the transcriptional activity of FoxM1, respectively. In addition, the relationship between anti- tumor effect of plumbagin and FoxM1 was investigated in vivo. Plumbagin significantly inhibited proliferation and induced apoptosis of esophageal squamous carcinoma cell in vitro and in vivo. Moreover, plumbagin down-regulated the expression of FoxM1 through suppression of its gene transcription. Our findings suggest that plumbagin may inhibit the proliferation of esophageal squamous carcinoma cell in vivo and in vitro through down-regulating the expression of FoxM1.
ABSTRACT
OBJECTIVE: To explore the inhibitory effect of betulinc acid(BA) on esophageal squamous cell carcinoma (ESCC) KYSE170 cells and the mechanisms of the inhibitory effect. METHODS: Different concentrations of BA(0, 5, 10, 20, 40, 60, 80 and 100 μg·mL-1) were used to detect their effects and the inhibitory rate on the cells for 24 to 72 h. The clone formation test was used to detect the long term inhibitory effects of different BA concentration(0, 5, 20 and 40 μg·mL-1) on KYSE170 cells. Different concentrations of BA(10, 60 and 100 (μg·mL-1) for 24 and 48 h were used to detect the apoptosis rate of cells by flow cytometry. Different concentrations of BA(0, 10 and 40 μg·mL-1) for 24 h were used to detect the cell cycle of cells by flow cytometry. RESULTS: BA inhibited the growth of KYSE170 cells in a dose- and time-depentent manner. IC50 at 24, 48 and 72 h were (56.81±2.56), (39.73±2.77) and (29.28±3.05) μg·mL-1, respectively. The clone formation plating efficiencies were (89.56±5.00)%, (61.00±2.03)%, (31.33±3.51)% and (15.33±2.33)% when the drug concentrations were 0, 5, 20 and 40 μg·mL-1, respectively. With the increase of BA concentration and the prolong of BA incubation time the apoptosis rate increased significantly (P<0.05 or 0.01). After 0, 1 and 10 μg·mL-1 BA added for 24 h, the rate of Gl phase cells decreased, while the rate of S phase cells increased(P<0.01). CONCLUSION: BA can inhibit the growth of KYSE170 cell by inducing cell apoptosis and blocking cells to stay in S phase. Copyright 2012 by the Chinese Pharmaceutical Association.