| Surgery resection is the only radical cure for cholangiocarcinoma,because of its hidden onset characteristics,Generally no obvious symptoms at the early late.Looking for natural antineoplastic drugs of high sensitivity and small side effects for Patients who cannot operate has great significance.Diosgenin,as a kind of widely used steroid compounds,known to possess anti-inflammatory oxidation resistance,anti-infectious activity,anticoagulation and thrombosis,and can be useful,for instance,in blood and cerebral disorders,allergic diseases,nerve protection,menopausal symptoms,and skin aging and desensitization,diabetes and obesity.Furthermore,it is an important starting materials of a variety of steroid hormone drugs clinically.In recent years,the anticancer effect of diosgenin has been reported in many different tumors,but few studies have been done on the specific role and mechanism of Cholangiocarcinoma.We are trying to explore the specific mechanisms of diosgenin on bile duct cancer cells in vitro and in vivo,six cholangiocarcinoma cells were chosen as the research objects.Cellular level:Diosgenin can inhibit Cholangiocarcinoma cell growth by the MTS with a concentration and time dependence,in addition,showed obvious inhibiting effects for cells clone formation.The Flow cytometry results showed that diosgenin could induce the G2/M period cell cycle arrest and apoptosis.WB showed that the protein expression of Cyclin B1 level decreased with drug concentration increased,and P21 were rising slightly,suggesting that diosgenin may through the regulation of P21 and Cyclin B1 proteins caused cell cycle arrest,then,blocked the cell cycle process leading to growth inhibition and apoptosis.We also found a significant rise in the proportion of Bax/Bcl-2,Caspase 3 and PARP-1 obvious fragmentation,cytochrome C from mitochondria released into the cytoplasm,z-VAD-FMK,Caspase broad-spectrum inhibitors,can be partially reversed apoptosis,From the above,diosgenin likely induced Cholangiocarcinoma cells apoptosis through the endogenous mitochondria pathway.In addition,diosgenin can inhibited the phosphorylation of the ninth serine of GSK3β,promoted the phosphorylation of the 216 tyrosine in GSK3p,increased the activity of GSK3β,and promoted the degradation of β-catenin.Thus diosgenin probably affected cholangiocarcinoma cells proliferation and induced apoptosis through regulating the GSK3 beta/beta-catenin signaling pathway.Animal level:We established the cholangiocarcinoma HuCCT1 cells tumor animal models in nude mice transplantation,the experiment shows that diosgenin could effectively inhibit cell growth in vivo with no obvious side effects.HE staining and WB results showed that diosgenin had the effect of inhibiting proliferation and promoting apoptosis in nude mice.The GSK3β/β-catenin signaling pathway was consistent with the cell level.The above results showed that diosgenin still had the ability to inhibit the proliferation and induce apoptosis of cholangiocarcinoma cell in vitro which was consistent with the cell level results.In conclusion,diosgenin has inhibitory effect on the proliferation of Cholangiocarcinoma cells. |
