Studies On Colon Cancer Prevention Effect And Its Mechanism Of Coptisine

Aim:Coptisine,one of the characteristic compositions of Rhizoma Coptidis,has been suggested in many studies to be an anti-oxidant,anti-inflammatory,antibacterial as well as a modulator of lipid and lipoprotein metabolism.The aim of the present study was to assess the cancer prevention effect of coptisine and elucidate its mechanism.Methods and Results:1.The colon cancer prevention model was established.All the mice were divided into five groups: normal control group,tumor control group,50 mg/kg Coptisine,100mg/kg Coptisine and 150 mg/kg Coptisine group.To induce the colon tumor,HCT-116 cells were subcutaneous injected in the right foreleg of nude mice.The nude mice with HCT-116 cells xenografts were treated with drugs via intragastric administration daily since two days before injection.At the time of sacrifice(day 25),the Coptisine groups had reduced in tumor weight and volume compared with tumor control group.Also,there were significant differences in levels of these projects between 150 mg/kg Coptisine group and tumor control group.Moreover,blood samples of all groups were collected for multiple tumor marker protein biochip assay.It suggested that levels of CEA,C A19-9 and CYFRA 21-1 had reduced significantly after treatment.During this period,although the body weight had decreased after coptisine treatment,there was no effect on the ratio between organ and body weight of nude mice,which means coptisine has no harmful effects on nude mice.These results confirm that coptisine has reasonable prevention of colon tumor in vivo.2.In this study,it was expected to excavate potential pharmacological activities of coptisine through analyzing potential protein targets.Targets of coptisine was evaluated and rapidly forecasted with reverse pharmacophore matching methods.The results were used to construct the d rug-pathways and “drug-targets-diseases” network.When the targets were classified into groups according to their related diseases,cancer was in the spotlight with 4 main targets(AKR1C3,CCNA2,ESR1 and IL2)and was effected though PI3K-Akt signaling pathway and pathways in cancer.Coptisine was also related to five other diseases,such as metabolic disease,cardiovascular disease and immune disease.3.In the vitro study,molecular mechanisms of coptisine induced HCT-116 growth and proliferation were studied.We focused mainly on roles of apoptosis and cell cycle arrest after coptisine treatment.The results of MTT and trypan blue assay expressed that coptisine could suppress tumor cells growth.The results of Annexin V/PI staining and Hoechst 33342 staining showed that this drug could induce apoptosis in HCT-116 cells.The effect of coptisine on the transcriptional and translational levels of key genes suggested that caspase-dependent apoptotic pathways were triggered in COP-treated HCT-116 cells.Additionally,we found that high dose coptisine exhibited significant inhibition of proliferation though causing the cell cycle arrest.When the cells were treated with coptisine for 24 h,G0/G1 phase cell cycle was arrested,levels of C yclin D1 and Cyclin E decreased significantly.4.The study found that coptisine significantly down-regulated the transcription of both oncogene KRAS and TNF-? in the tumor tissues by MAPK pathway,as well as up-regulated the expression of tumor-suppressor genes p53.Similarly,coptisine significantly down-regulated the transcription of K RAS,BRAF and MEK-ERK pathway in the lung tissues.Conclusion:The present study proved that coptisine has colon cancer prevention effect.It has been shown that coptisine prevented cell proliferation in vivo,and induced apoptosis in HCT-116 cells.These data suggest that coptisine deserves further studies as a colon cancer prevention agent in clinical trials.