Mechanism Of Raddeanin A (RA) Against Gastric Cancer Based On Autophagy And Apoptosis

Research Purpose:Raddeanin A (RA) is an extractive from Anemone raddeana Regel, a traditional Chinese medicine. Our previous study showed that RA can induce apoptosis and inhibit the invasive function of gastric cancer (GC) cells function. This research intends to further explore the mechanism of apoptosis. In addition, the theoretical study, we found that autophagy is also one of the anti-tumor mechanisms. Thus, this study was to investigate its autophagy of GC cellss and its mechanism, and to further explore the relationship between RA induced autophagy and apoptosis.Research Methods:1 MTT assay was used to detect RA inhibition of GC cells (SGC-7901, SNU-1, HGC-27) proliferation.2 Flow cytometry analysis and Hoechst 33258 staining was used to observe the effects of RA on apoptosis of GC cells.3 Transmission electron microscopy assay was used to observe RA-induced GC cells autophagy formation.4 Western blotting was used to observe the changes of expression of related proteins.5 We suppressed or excited autophagy. Then, we observed the effect of the RA-induced GC cells proliferation, apoptosis, and autophagy.Research Results:1 MTT assay showed that RA inhibition of proliferation of GC cells increased with a dose-dependent manner.2 Flow cytometry analysis and Hoechst 33258 staining showed that RA induced apoptosis on GC cells.3 Transmission electron microscopy assay found that RA can induce the formation of GC cells autophagy.4 Western blotting showed that the RA induces apoptosis and autophagy by activating p38 MAPK pathway, and inhibiting mTOR pathway.5 After the inhibition of autophagy, Flow cytometry analysis showed that apoptosis induced by RA was to be further strengthened. Western blottingting experiments confirmed these changes.Research Conclusions:1 RA activates p38 MAPK signal pathway and improv BAX/BCL-2 ratio to induce three strains of varying degrees of differentiation of GC cells apoptosis.2 RA activates p38 MAPK signal pathway, inhibits the mTOR pathway, induced three GC cells autophagy.3 Autophagy protects from RA-induced apoptosis in GC cells.