| Part 1 The expression and the mechanism of P-gp, MRP, LRP in the T24/ADM cellsObjective:Detection of MDR-1, MRP1, LRP mRNA in the T24/ADM cells and to explore its mechanism.Methods:concentrations of ADM was lowly stepwise increased for long explsure on T24 cells,to establish of drug resistant T24/ADM cell lines.Regard T24 cell lines as control group. MTT assays of multidrug-resistant T24/ADM cells sensitivity to drug, Observing the distribution of ADM in the T24/ADM cells and the effect of verapamil(antagony P-gp) on it by Confocal microscope. Detecting the mRNA expression level of multidrug resistance gene lung resistance-related protein gene and multidrug resistance-associated protein gene by RT-PCRRusults:the drug resistanc of T24/ADM cell reinforced obviously,its resistance index of T24/ADM cells to ADM was 4.86. The mean fluorescence of ADM in T24/ADM cell cytoplasm was 504.380±79.5 and in nuclei was 166.304±18.6, both of them are lower than T24 cell's (715.058,226.807), p< 0.05. after Ver intervention, the mean fluorescence of ADM In T24/ADM cytoplasm was 624.097±60.5, higher than it (504.3802±79.5) was not concerned by Ver, p<0.05. RT-PCR detected that,the Relative expression of MDR-1 mRNA and MRP1 mRNA in T24/ADM was 0.382±0.075 and 1.053±0.081, higher than it expresses in T24 cell, p<0.05. The Relative expression of LRP mRNA was 0.792±0.068 in T24 cell, it was 0.823±0.052 inT24/ADM, They are not difference to each other, p>0.05。Conclusion:①The overexpression of MDR-1 gene lead the concentration of drug in cell to degrede,and then lead T24/ADM bladder cancer to resist drug. It is a molecular basis for the T24/ADM cell.②The expression of MRP 1 gene step up in T24/ADM cell obviously,but we don't know its mechanism of action clearly, we need to study it further.③The overexpression of LRP gene in T24 and T24/ADM cells are not very difference,it is't a MDR molecular basis for the T24/ADM cell.Part 2 The expression and significance of MDR-1, MRP1, LRP in situ nude mice T24/ADM bladder carcinomaObjective:Detection of MDR-1, MRP1, LRP mRNA in T24/ADM cells and to explore its importance. Methods:Injecting T24/ADM and T24 cells into two groups of nude mice,to found situ nude mice T24/ADM bladder carcinoma model and situ nude mice T24 bladder carcinoma model.their tumor tissue were collected,to detected the MDR-1, MRP1 and LRP gene mRNA expression levels by RT-PCR,and to made Tumor cell suspension by enzyme digestion, for detecting their resistance to antitumor drug by MTT assay,and for observing the distribution of ADM in their cytoplasm and nuclei by confocal microscopic.Rusults:the Relative expression of MDR-1 mRNA and MRP1 mRNA in T24/ADM tumor tissue was 0.382±0.075 and 1.053±0.081, higher than them expresse in T24 tumor tissue, p<0.05. The Relative expression level of LRP mRNA was 0.813±0.065 in T24/ADM tumor tissue, it was 0.725±0.051 in T24 tumor tissue, They are not difference to each other, p>0.05。The results of MTT:the drug resistanc of cells of T24/ADM tumor tissue reinforced obviously,and its resistance index to ADM was 3.30.Confocal microscopy observations: The mean fluorescence of ADM in T24/ADM tumor tissue cell cytoplasm was 503.185±44.6 and in nuclei was 185.426±19.0, both of them are lower than T24 tumor tissue cell(725.530±49.1, 218.589±24.3), p<0.05.Conclusion:①MDR-1 is overexpressive in the multidrug resistance situ bladder carcinoma model of nude mice.②The MRP1 gene still overexpress in the multidrug resistance situ bladder carcinoma model.③the expression of LRP gene is not very different in T24/ADM and T24 situ bladder carcinoma model of nude mice.can't be a MDR molecular basis for the the multidrug resistance situ bladder carcinoma model.
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