The Effect Of Membrane Toxin-12 On The Biological Behaviors Of Bladder Cancer Cell Lines

OBJECTIVE:In this study,we explored the effects of membrane toxin(MT-12)of Chinese cobra on the proliferation,migration,adhesion and apoptosis of bladder cancer cell lines(RT4 and T24)in vitro.METHODS:1.After bladder cancer RT4,T24 cell lines were treated by various concentration of MT-12(0,0.25 and 0.5μg/mL)for 0,1,2,3,4,5d,the ability of the cell proliferation in bladder cancer cell lines was measured by CCK8 assay.Cells in control group were treated by PSA.2.After bladder cancer RT4,T24 cell lines were treated by two concentration of MT-12(0 and 0.5μg/mL)for 6,24h,the ability of the cell apoptosis in bladder cancer cell lines was measured by Annexin V-FITC/PI staining followed flow cytometry,in which the amount of the apoptosis rate was used to measure the apoptosis ability.3.Wound healing cell migration assay was used to determine the ability of cell metastasis in bladder cancer RT4,T24 cell lines that were treated by various concentration of MT-12(0,0.25 and 0.5μg/mL)for 0,24h.The cell migration distance is determined by measuring the width of the wound.4.After bladder cancer RT4,T24 cell lines were treated by various concentration of MT-12(0,0.25 and 0.5μg/mL)for 24h,the ability of cell adhesion was determined by MTT assay,in which the amount of cells which remain in plates was used to measure the adhesion ability.RESULTS:1.MT-12 inhibited bladder cancer cell lines(RT4 and T24)on growth in time-concentration-dependent manners showing the phenomenon that with the increase of the concentration and the prolongation of the action time the inhibition effect on cells is stronger.2.MT-12 inhibited bladder cancer cell lines(RT4 and T24)on apoptosis in time-dependent manners showing the phenomenon that the inhibition effect on cells is stronger with the prolongation of action time.3.MT-12 inhibited bladder cancer cell lines(RT4 and T24)on migration,adhesion in concentration-dependent manners showing the phenomenon that with the increase of the concentration the inhibition effect on cells is stronger..CONCLUSION:MT-12 inhibited bladder cancer cell lines(RT4 and T24)on growth in time-concentration-dependent manners,inhibited the migration,adhesion of bladder cancer cell lines(RT4 and T24)in concentration-dependent manners.And inhibite the apoptosis in bladder cancer RT4 and T24 cells in a time-dependent mamnner.The study proves that MT-12 produces cell death through the endogenous pathway of apoptosis.