Study On The Effect Of Demethylzeylasteral To Bladder Cancer Cell Line T-24

Background:Bladder cancer is the most common malignant tumor in urology system. the postoperative recurrence rate is very high,while retaining the bladder after surgery for about 50%-70%of patients with postoperative recurrence.Urology by scholars at home and abroad over the years to explore,discover Intravesical chemotherapy drugs regularly can be effective in reducing and delaying the recurrence of bladder tumor,improve the survival rate of patients.Therefore,the use of chemical treatment before and after surgery,as well as the use of advanced bladder cancer chemotherapy are the current treatment of bladder cancer treatment and the focus of the study.Choose the appropriate chemotherapy is the current treatment of bladder cancer hot spots.In recent years,the role of anti-tumor Tripterygium wilfordii has been more and more attention,but because of the complex components of Tripterygium wilfordii people eager to be separated from the monomer composition effective to find an ideal anti-tumor monomer.The topics on the latest extract of Tripterygium wilfordii demethylzeylasteral(Dzy,T-96) on the role of bladder cancer study.Part OneStudy on the toxic effect of demethylzeylasteral to bladder cancer cell line T-24Objective:Investigate the toxic effect of demethylzeylasteral to bladder cancer cell line T-24.Select certain concentration of demethylzeylasteral for the next studies.Methods:Cytoactivaty of T-24 cells were measured after treated with varies concentrations of THP and demethylzeylasteral with the method of MTT.The index of cytotoxicity(IC) was calculated.Results:IC of THP and demethylzeylasteral were increased with the increase of the concentration.A " flat phase" turns up after a certain concentration both in the group of THP(lug/ml).Conclusion:Take the pharmacokinetics account,two different concentrations of each drug were selected for the next studies.Part TwoStudy on the cytotoxicity of demethylzeylasteral to bladder cancer cell line T-24Objective:Investigate the dose-effect and time-effect connection of THP and demethylzeylasteral to bladder cancer cell line T-24.Whether there is a significant difference between the cytotoxicity of THP and demethylzeylasteral.Methods: Cytoactivity of T-24 cells were measured after treated with varies concentrations index of cytotoxicity(IC) was calculated,too.Results:IC of THP and demethylzeylasteral were increased with the increase of treated time.There is a significant difference between IC of the lower and higher concentration group of THP and demethylzeylasteral on the third day(lower group:0.624±0.035 vs 0.563±0.032,P=0.015;higher group:0.792±0.026 vs 0.672±0.028,P＜0.001 ).There is no significant difference between IC of the lower and higher concentration group of THP and demethylzeylasteral on the first and second day(Lower group:the first day:0.152±0.063 vs 0.127±0.055,P=0.456,the second day 0.504±0.031 vs 0.507±0.042,P=0.879;Higher group:the first day:0.266±0.068 vs 0.231±0.057, P=0.256,the second day 0.585±0.053 vs 0.594±0.039,P=0.660).Conclusion: Whether the lower or higher concentration groups of THP and demethylzeylasteral have a similar cytotoxicity to bladder cancer cell line T-24.Part ThreeThrough Annexin V and PI double staining,using flow cytometry to study the way of demethylzeylasteral effect on bladder cancer ceU line T-24Objective:study the way of demethylzeylasteral effect on bladder cancer cell line T-24(to kill cells or induce apoptosis),explore the possible mechanisms of demethylzeylasteral effect on tumor cells.Methods:Through Annexin V and PI double staining After treated with THP and demethylzeylasteral.Then,screened by flow cytometry to distinguish among them were killed or apoptosis of cells.Result: Demethylzeylasteral:Lower group:Necrotic cells8.29%±1.03,Apoptotic cells 14.37%±2.50;Higher group:Necrotic cells 27.935%±3.80,Apoptotic cells 55.105%±9.75;THP:Lower group:Necrotic cells 64.41%±15.53,Apoptotic cells 2.16%±1.91;Higher group:Necrotic cells 96.25%±2.28,Apoptotic cells 2.81%±2.49 Conclusion:demethylzeylasteral mainly by inducing cell apoptosis pathway effect on bladder cancer cell line T-24.Part FourStudy on the relationship of demethylzeylasteral with bax and caspase-3 gene in inducing bladder cancer cell line T-24 apoptosisObjective:study on the relationship of demethylzeylasteral with bax and caspase-3 gene in inducing bladder cancer cell line T-24 apoptosis.Methods:Use Immunohistochemical method,detect qualitatively the expression of bax and caspase-3,after treated with demethylzeylasteral.Compare with treated with THP and nothing.Result:bax and caspase-3 expression was positive in T-24 cell after treated with demethylzeylasteral.Bax and caspase-3 expression were positive after treated with THP.Conclusion:demethylzeylasteral induce T-24 cell apoptosis, Activate bax and caspase-3 gene expression may be possible mechanisms.