The Research Of The Mechanism Of Survivin In The Process Of The Human T24Bladder Cancer Cell’s Chemotherapy Apoptosis

Objective: Survivin is a new member of the family of the inhibition of apoptosisprotein (IAP) which was found in recent years, it is that the strongest apoptosisinhibiting factor as far, and survivin is most widely high expression in humanmalignant tumor tissues, rarely expressed in normal tissue or not. Studies have shownthat the factor "survivin" plays an important role in the formation and developmentof tumor by the way of inhibiting tumor cell apoptosis. This study detections theproliferation of the human bladder cancer T24cell, and detections the expressinglevel of survivin mRNA and survivin protein in the human bladder cancer T24cellunder the action of different concentration of chemotherapy drugs Pirarubicin(THP).To discusses the mechanisms and principles of survivin in the process ofthe human bladder cancer T24cells ’apoptosis by THP. And this study may providecertain theoretical basis for the clinical infusion treatment of THP of the humanbladder cancer, it also can provide certain theoretical basis for the occurrence anddevelopment and the molecular target therapy of the human bladder cancer.Methods:1. configured to different final concentration of THP (0,0.1,1,10ug/ml), respectively to act on human bladder cancer T24cells in a certain time, in setup blank control group and drug group under the premise,the MTT method wasapplied to detect the inhibition rate of proliferation under the different concentrationof the human bladder cancer T24cells after the effection of THP respectively24, 48,72hours.2.The RT-PCR method was applied to detect the survivin mRNAexpression level in different concentrations (0,0.1,1,10ug/ml) group of T24bladdercancer cell after the action of the THP for24hours.3. The application of Western blotprotein imprinting technology was to detect the expression level of survivin proteinin human T24bladder cancer cells in different concentrations (0,0.1,1,10ug/ml)THP action after24hours. Application of SPSS13.0statistical software for statisticalanalysis, all the datas were shown by the way of "means-standard"(X+s). Twosample mean is using the T test, multiple mean to compare this adopt completelyrandom single factor analysis of variance (One-one-way ANOVA). P <0.05showedthe difference of the different group was statistically significant, P <0.01showed thedifference was statistically significant greatly.Results:1.Each group OD value was detected by MTT method included theblank group,24h,48h,72h the four groups of T24cells after the THP action,according to the measured OD value, to compare each cell’s growth situation.Compared with control group, each group concentration of THP in24h,48h,72h allcan inhibit the T24bladder cancer cell proliferation, which has statistically significantdifference (P <0.05), calculated the proliferation inhibition rate of the drug groupafter24h,48h,72h,it was found its inhibition rate present certain concentration andtime dependence (P <0.05).2. After24hours action of different concentrations (0,0.1,1,10mu g/ml) THP,the survivin mRNA expression of each group T24cells allmake significantly declined, and performance for certain concentration dependence(P <0.01).3. The application of western blot was tested the survivin protein in eachgroup of different concentrations of THP after24hours, the expression of survivinprotein of each group significantly declined. With the increase of the THPconcentration, the more obvious of the cutting down of survivin protein showed acertain range of concentration dependence (P <0.05).Conclusions: In the vitro experiments of T24cells, the THP has obvious inhibitory effect on T24bladder cancer cells, and the effection has a certainconcentration and time dependence. That mean with the increase of the concentrationof THP and the extension of the action time, the THP inhibit the T24cell’s growthmore apparently. In the vitro experiment of THP, the survivin mRNA and protein asthe enhancement of the action of THP on T24cell, namely the increase ofconcentration of THP and the extension of action time, the expression of the survivinmRNA and protein decreased more obviously, which above showed a certainconcentration and time dependent. Above all, THP is likely to be cut the expression ofsurvivin’s apoptosis inhibiting effection to push theT24cells, apoptosis which throughthis pathway.And the THP was mostly like to be in the principle of above said downto inhibit the growth of T24bladder cancer cell.