Study On Apoptosis Effect Of Bladder Cancer Cell Line T₂₄ Induced By BCG Polysaecharide And Nucleis Acid Combined With Cisplatin

Background:BCG polysaccharide nucleic acid（BCG-PSN）is an immunologic active material manufactured after the mixing of the extraction of the polysaccharide and nucleic acid in the BCG with 65℃ hot phenol alcohol method,it has the immunoregulation,antitumor,anti-inflammatory,antivirus and anti-allergic effects.Cisplatin（DDP）and its derivatives are the most common and effective drug in treating tumors clinically so far.However,the latest statistical data shows that during the chemotherapy process,the use of DDP clinically is restricted greatly no matter the intrinsic or the acquired drug resistance.Therefore at present,what we are pursuing is changing the tumor’s multidrug resistance to improve the efficacy of chemotherapy drugs.Objective:The experiment objective is to study DDP’s combination with different concentration of BCG-PSN to the influence of the multiplication,apoptosis,invasion and migration abilities of the human carcinoma of urinary bladder cell line T₂₄.Methods:1.The experiment is divided into blank group（no drug group）,DDP（0.25ug/ml）,DDP adding different concentration of BCG-PSN group（0.25ug/ml+0.05mg/ml,0.25ug/ml+0.15mg/ml,0.25ug/ml+0.25mg/ml）.2.Apply CCK-8 to observe after DDP adding different concentration BCG-PSN in different action period（12,24h）,T₂₄ cell’s inhibition.3.Flow cytometer detects DDP adding different concentration of BCG-PSN to intervene T₂₄ apoptosis.4.Artificial reconstituted basement membrane test to Transwell,check DDP adding different concentration of BCG-PSN’s influence to T₂₄ cell invasion ability.5.Scratch healing test to check DDP adding different concentration of BCG-PSN’s influence to T₂₄ cell migration ability.Results:1.CCK-8 test result shows: after DDP’s combination with different concentration of BCG-PSN and taking effect on T₂₄ cell for 12 hours or 24 hours,it has significant inhibition（P < 0.05）comparing to the blank group,and presenting BCG-PSN action concentration and time dependency.Comparing to Cisplatin,DDP group,the inhibition ratio of DDP+BCG-PSN（0.05mg/ml）group is 38.6% after taking effect on cell for 12 hours,without any statistical difference（P>0.05）.But its inhibition ratio significantly improves after 24 hours,is statistically significant（P<0.05）.For the rest drug combination group,the multiplication inhibition ratio is significantly improved（P<0.05）.2.Flow cytometer detection analysis discovers that after DDP’s combination with different concentration of BCG-PSN and taking effect on T₂₄ cell for 12 hours,the number of apoptotic cell presenting increased dependence of BCG-PSN action concentration.The total apoptotic cell range is 7.03%-59.84%.3.The invasion experiment shows that after DDP’s combination with different concentration of BCG-PSN and taking effect on T₂₄ cell for 24 hours,T₂₄ cell invasion ability is significantly lowered with the BCG-PSN concentration increase.4.The migration experiment shows that after DDP’s combination with different concentration of BCG-PCN and taking effect on different times（0,6,12 and24hours）,T₂₄ cell’s migration ability is gradually lowered,which presents BCG-PSN action concentration and time dependency.Conclusions:BCG-PSN combining with Cisplatin,DDP can significantly inhibit the multiplication of human bladder cancer cell line T₂₄,quicken apoptosis,lower its invasion and migration abilities,and presents BCG-PSN significant dose dependency.