Research Study Of Toosendanin For Colorectal Cancer SW480Cell Inhibition And It’s Mechanism

Objective:This study aims to explore toosendanin’s inhibition effect on colorectal cancer cell SW480and the relevant molecular mechanism.Method:MTT method was used to observe the effects of different concentrations of toosendanin on human colorectal cancer cell line SW480proliferation and the relationship between drug effect and time; Flow cytometry, AnnexinV/PI double staining method was used to observe the effect of toosendanin on human colorectal cancer cell line SW480apoptosis; Fluorescence microscope, Hoechst33342staining method was used to observe the morphology changes of human colorectal cancer cell line SW480apoptosis; Flow sytometry was used to detect toosendanin’s effects on the cell cycle of human colorectal cancer cell line SW480; Confocal laser scanning microscope was used to observe the nucleus translocation of β-catenin after toosendanin treatment on human colorectal cancer cell line SW480;Real-time PCR method to detect apoptosis-gene expression after toosendanin treatment on colorectal cancer SW480; Western Blotting method to detect apoptosis-protein expression after toosendanin treatment on colorectal cancer cell line SW480. Resμlts:0.031μM-0.5μM concentration of toosendanin inhibited the proliferation of human colorectal cancer cell line SW480with concentration and time-dependence.0.125μM and0.5μM concentration of toosendanin treatment on human colorectal cancer cell line SW480for48h induces early apoptosis in a concentration-dependent manner, and morphological changes as apoptic bodies are found; compared with negative control group, when human colorectal cancer cell line SW480were treated with0.125μM,0.25μM and0.5μM concentration of toosendanin for24h, as the dose concentration increases, hyodiplod cells are detected before G0/G1phase, the percentage of hyodiplod cells increases from19.40%to23.0%at S phase, and the cells are arrested in S phase.0.5μM concentration of toosendanin treatment on human colorectal cancer cell line SW480for48h, β-catenin transfers to the outside of nucleus.0.125μM and0.5p.M concentration of toosendanin treatment on human colorectal cancer cell line SW480for48h up-regulates Bax,Bak and GSK-2β, but down-regulates Bcl-2,Bcl-xL,Survivin,cyclin Dl,cyclin D2,cyclin D3,β-catenin,VEGFA,c-myc and COX-2.0.125μM and0.5μM concentration of toosendanin treatment on human colorectal cancer cell line SW480for48h up-regulates protein Bax and GSK-3β,down-regulate protein Bcl-2,Pro-caspase3,Pro-PARP,P-AKT Ser473,AKT,P-GSK-3β Ser9,β-catenin and VEGFA.Conclusions:Toosendanin inhibits the proliferation of human colorectal cancer cell line SW480;Toosendanin can induce human colorectal cancer cell line SW480apoptosis;Toosendanin’s inhibition on human colorectal cancer cell line SW480can possibly be realized through AKT/GSK-3β/β-catenin signaling pathway.