Cyclosporin A induced apoptosis in human gastric adenocarcinoma MGC80-3 cells and Inhibitory effect on P-gp expression

Aim To study cyclosporin A(CsA) induced apoptosis in human gastric adenocarcinoma MGC80-3 cells and the effect on P-gp expression and further explore the relationship between them.Method The proliferation inhibition of human gastric adenocarcinoma MGC80-3 cells was determined by MTT assay after different concentration of CsA treatment at 24,48 and 72 h.Annexin V-FITC / PI double staining was used to detect the cell apoptosis produced by CsA after 48 h,and acridine orange / ethidium bromide(AO / EB)fluorescent staining was utilized to observe morphological changes of apoptotic cells.Western blot was applied to examine the expression of P-gp in MGC80-3 cells.Results CsA suppressed human gastric adenocarcinoma MGC80-3 cells growth and proliferation in a dose-dependent manner within the concentration ranging from 0 to 25.0 μmol·L-1 and in a time-dependent manner from 24 h to 72 h.After treated for 48 h with different concentration of CsA,the cell shape was irregular and typical morphological changes of apoptosis such as nuclear chromatin condensation were also evaluated by AO / EB fluorescence staining test.FCM analysis showed that CsA-induced apoptosis against MGC80-3 cells was in a concentration-dependent manner and the cell apoptosis had significant difference compared with the control group(P 0.05 or P 0.01).Western blot showed that CsA also significantly reduced the expression levels of P-gp in a concentration-dependent manner(P 0.05 or P 0.01).Conclusion CsA may induce apoptosis and down-regulate the expression levels of P-gp in human gastric adenocarcinoma MGC80-3 cells,and the inhibitory effect on P-gp may be related to apoptosis.