The Primary Experimental Study Of The Inhibition On Bladder Cancer T24 Cell Line With Composition Of Drugs

ObjectiveThe bladder cancer is the common malignancy of the genitourinary system in Chinese.The superficial bladder cancer(non-muscular invasive bladder cancer) accounts for about 75% of the bladder cancer. The major treatment is transurethral resection of bladder tumor (TURBt).The highest recurrence rate is 67% after TURBt. The adjuvant intravesical chemotherapy following TURBt must be used in all patients and may reduce future recurrences except about 10-30% of its recurrences. It is to seeking the effective intravesical instillation drugs to reduce recurrence rate that is important in bladder cancer treatment.Mitomycin C(MMC),hydroxycam-ptothecine(HPT),epirubcin(EPI) and Adriamycin(ADM) are selected generally in clinical treatment.It is not exact that the effect of any drug is best. It was reported that combined chemotherapy was better than single drug in reduce the inhibition ratio of bladder cancer.It is not exact that the effect of any drug combined chemotherapy is best.Rapamycin(RAPA) is a immuno-suppressant antibiotics and play an important role in the anti-tumor, so it is used in patients who receiving renal transplantation with transitional cell carcinoma that is a common carcinoma among it most is bladder cancer.It was reported that there is not using RAPA in patients who is suffering from bladder cancer and not receiving renal transplantation.MMC,RAPA,HPT and EPI were used in this study.To obtain the largest inhibition ratio of four drugs alone or in combination with each other on bladder cancer T24 cell line in vitro and study primary mechanism on effect of drugs.To provide the theoretical basis for clinical drug using to prevent recurrences following TURBt henceforth.Methods To obtain the best consistency of bladder cancer T24 cells in vitro.By cells culturing in vitro,MTT and some formulas were used to obtain the IC50 of the four drugs.T24 cells were incubated with the IC50 of the four drugs alone and in combination with each other(all and half of IC50) to obtain the largest inhibition ratio.The experiment is timed two periods that 24 and 48 hours(after treated with drugs for 24 hours and 48 hours). Then the cell cycle was analyzed by the FCM with PI and apoptosis was observed by the microscope with HE in each group.SPSS was used in statistical analysis with analysis of variance and SNK-q test.ResultThe experimental consistency of bladder cancer T24 cells were 10×104/ml.The IC50 of MMC,HPT,RAPA,EPI were 12μg/ml,5μg/ml,125ng/ml, 11μg/ml(the dose of the experiment). (1)The inhibition ratio of four drugs MMC,HPT,RAPA,EPIwere 35.89%,12.92%,6.70%,49.28% for 24 hours.The inhibition ratio of EPI was significantly higher than other three drugs for 24 hours that the difference was statistical significance. (2)The inhibition ratio of four drugs MMC,HPT,RAPA,EPI were 86.08%,72.16%,41.39%,87.91% for 48 hours. The inhibition ratio of EPI,MMC,HPT whose difference were statistical significance compared with RAPA,and there were not statistical significance among EPI,MMC,HPT.(3)The largest inhibition ratio of every group of four drugs and its composition were the composition of MMC and RAPA(half and all of IC50)whose inhibition ratio were73.68%,63.64%for 24 hours that the difference were not statistical significance.That the inhibition ratio of the composition larger than the EPI and the difference was statistical significance.(4)The largest inhibition ratio in all drugs and its composition is the composition of MMC and RAPA(all of IC50) that it was 98.53% for 48 hours.That the inhibition ratio of the composition of MMC and RAPA(all of IC50)was higher than RAPA and the composition of RAPA and HPT(half of IC50) whose inhibition ratio were 41.39%,60.73% that the difference was statistical significance.The lowest inhibition ratio in all drugs and its composition is RAPA.That the inhibition ratio were no difference among the drugs and its composition except its.The result of FCM indicated that the cell cycle at G1 phase of MMC,RAPA and its composition(all and half of IC50) were 65.46±0.16%,69.36±0.35%,53.37±0.51%,59.02±0.42%.The cell cycle at G1 phase of RAPA was higher than others.That the difference was statistical significance.It was observed by microscope with HE that a number of apoptosis after treated with drugs.ConclusionThe largest inhibition ratio in four drugs is the EPI among MMC,HPT,EPI,RAPA.The largest inhibition ratio in every group of four drugs composition was the composition of MMC and RAPA that was higher than the four drugs.Using RAPA in combined chemotherapy on bladder cancer cell bring forth new ideas and lay foundations for using RAPA in patients who is suffering from bladder cancer and not receiving renal transplantation.