Improvement of Treatment for Urological Cancer by controlling Function of Apotosis-associated molecules using siRNA.

1. XIAP被siRNA或shRNA敲除。设计并引入XIAP siRNA或shRNA，分别瞬时或连续降低XIAP的产量。使用DU145建立前列腺癌模型(1)抗肿瘤药物敏感性和敲低XIAP表达。短暂抑制XIAP导致细胞对CDDP的敏感性增加，但持续抑制诱导的抗性高于亲本细胞。BCL-2和NF κ-B在稳定的转染中表达上调。(2)通过抗bcl -2小分子HA14-1恢复敏感性。HA14-1消除了cddp的易感性。利用Caki-1建立肾细胞癌模型。(1)正常肾脏和肾细胞癌（RCC）中XIAP和smac的表达。在正常肾中，肾小管细胞表达smac和XIAP。在34例rcc中，smac表达频率较低，XIAP在高分级和分期肿瘤中强烈表达。XIAP和smac分别在caki和KRC/Y中高表达。(2) HA14-1或smac-模拟合成肽（smac-ant）对Caki-1 XIAP低表达克隆细胞生长的影响。单独使用HA14-1对细胞生长有一定的抑制作用，但与抗fas抗体联合使用cddp和smacant治疗对细胞生长有较高的抑制作用。总之，XIAP似乎降低了前列腺和肾细胞癌对凋亡刺激的易感性。使用shRNA抑制可能不足以克服这种特性，而额外治疗降低BCL-2功能或使用smac-ant进一步抑制XIAP可能是有用的。

1. XIAP knock down by siRNA or shRNA.Designed and introduction of XIAP siRNA or shRNA, decreased production of XIAP transiently or continuously, respectively.2. Prostate cancer model using DU145(1) Sensitivity to anticancer drug and knock down of XIAP expression.Transient suppression of XIAP resulted in increase susceptibility to CDDP, but continuous suppression induced resistant character than parent cell. Upregulation of BCL-2 and NF κ-B were observed in stable transfectants.(2) Restore the sensitivity by anti-BCL-2 small molecule HA14-1.HA14-1 cancelled decrease the susceptibility to CDDP.3. Renal cell cancer model using Caki-1.(1) XIAP and smac expression in normal kidney and renal cell cancer (RCC).In normal kidney, smac and XIAP were expressed in renal tubular cells. In 34 RCCs, smac was expressed less frequently, XIAP was strongly expressed in high grade and stage tumor. XIAP and smac were highly expressed in Caki-I and KRC/Y, respectively.(2) Effect of HA14-1 or smac-mimic synthetic peptide (smac-ant) on cell growth in Caki-1 XIAP lower expresser clone.HA14-1 alone revealed modest suppression of cell growth but combination with anti-Fas antibody resulted in additional effect-CDDP and smac-ant treatment showed high suppression.4. In conclusion, XIAP seems to decrease susceptibility to apoptotic stimuli in prostate and renal cell cancer. Suppression using shRNA may not be enough to overcome such character, and additional treatment for reduction of BCL-2 function or further suppression of XIAP using smac-ant might be useful.