| Objective:This study investigated the effects of Andro on human gallbladder cancer and uncovered the molecular mechanisms responsible for these effects.This study could provide experimental evidence for the application of Andro as a new natural antitumor medicine for gallbladder carcinoma(GBC).Methods:In order to verify the effects of Andro on human gallbladder cancer in vitro several experiments had been down such as MTT assay,colony formation assay,flow cytometric analysis,hoechst 33342 staining,cell scratch and transwell migration assay.Moreover we tested the in vivo activity of Andro using xenograft modal of NOZ cells.We also explored the effect of Andro on the mechanism of gallbladder.Western blot analysis,qRT-PCR and immune histochemical staining were used to explore the molecular mechanism of andrographolide in gallbladder cancer cells in vivo and in vitro.Results:The resules showed that Andro could suppress proliferation of gallbladder cancer cells(GBC-SD and NOZ cells)in a dose-and time-dependent manner.With the increase of concentration of Andro,the cell apoptosis rate would be significantly increased and showed nuclear condensation apoptotic morphological changes.Results showed that Andro treatment down-regulated Bcl-2,up-regulated Bax,induced mitochondrial cytochrome C to the cytoplasmic release and induced mitochondrial cytochrome C to the cytoplasmic release,activatied of downstream caspase-9,caspase-3,induced mitochondria-mediated apoptosis.Andro significantly inhibited cell cycle progression in gallbladder cells with an obvious increase in the proportion of cells in S-phase.We also observed that the levels of cell cycle-related proteins decreased,showing that Andro treatment down-regulated cyclin D1 and cyclin A.Furthermore,Andro inhibited the growth of xenograft tumors in nude mice.We also explored the effect of Andro on the mechanism of gallbladder.We also found that Andro inhibited JAK2-STAT3 signaling pathway in vitro and vivo.Conclusion:Our findings reveal a novel drug action by Andro in gallbladder cancer cells and suggest that Andro can induce GBC-SD and NOZ cell mitochondria-mediated apoptosis and S-phase arrest and inhibit cell migration.Furthermore,Andro inhibited the growth of xenograft tumors in nude mice.Moreover,we discovered that the anti-cancer effects of Andro on gallbladder cancer may through or in part through the JAK2-STAT3 pathway.Therefore,Andro has potential as a novel anti-tumor therapy for the treatment of GBC. |
