The Study Of Genistein On ADM-Resistant Bladder Tumor Cells

Bladder tumor is urogenital system frequent malignant tumor.At present,therapeutic tools of bladder tumor are the operation,chemotherapy,radiotherapy and immunization adjunctive therapy.After the operation,making chemistries drug pour into the bladde is important method to cure bladder tumor. However,after the chemotherapy,bladder tumor has 40%～80%recurrence rate.Multi-drug resistance (MDR) remains one of the major obstacles to successful chemotherapy.More than 90%of tumor patients died of MDR.Consequently,there is still an urgent need of a valuable strategy to overcome MDR.In recent years,It is imperative to establish a MDR human bladder cancer cells line and explore therapy durg.Genistein is a kind of phytoestrogens,A growing of research has revealed that genistein decrease human breast cancer,human ovarian cancer,human prostate cancer etal.And partly revesed tumor cells,induced apoptosis,elevated chemotherapy effect.In this study,we aimed to observe effect of Genistein on multidrug-resistant cell lines T24/ADM.and to investigate possible mechanism of Genistein -induced MDR.PART ONE Establishment and Characterization of Multi-drug Resistant Bladder Tumor T24 Cell linesObjective:To establish multi-drug resistant bladder tumor T24 cell lines and to assess their resistant characteristics.Methods:Bladder tumor T24 cell lines was exposed to adriamycin(ADM) in the culture medium for the establishment of drug resistant cell lines:concentrations of ADM was stepwise increased for long exposure.Morphologic studies were performed with optical microscopy.Drug sensitivities were determined by MTT assay,MDR1mRNA were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Results:Six months were taken to establish drug resistant cell linesT24/ADM.No obvious morphologic changes were observed between resistant and parental cell.But,drug resistance to ADM,5-FU,CXT and CDDP heighten.RT-PCR showed that the expressions of MDR1 recreased remarkably.P＜0.01 among all groups there are significant differences.Conclusion.Cell lines T24/ADM were multi-drug resistant,and it's drug resistance related to the overexpression of mdr-1 gene production P-gp.It is qualified to further research on multidrug reversing. PART TWO Proliferation Induced by Genistein In Multi-drug Resistance Bladder Tumor T24 CellsObjective:To observe Effect of Genistein on multidrug-resistant cell lines T24/ADM.Methods:Use Genistein to irradiate T24/ADM cell.The sensitivity of T24/ADM cells to genistein was detected by MTT assay.Results:After Genistein was used to T24/ADM cells,The IC₅₀ value of genistein was 55mg/L.The apoptotic cells were induced by genistein at the concentration of 4-100mg/L.Conclusion:Genistein can inhibit human urinary bladder cancer T24/ADM cell and induce apoptosis at some concentration.The induction of apoptosis is a very important mechanism of genistein to treat urinary bladder cancer.