【摘要】 目的:探讨Survivin对子宫内膜癌细胞运动、侵袭能力的调节作用及其可能机制。方法:将含有Survivin特异性siRNA的重组质粒pRNAT-suv和阴性对照质粒pR-NAT-neg分别转染子宫内膜癌细胞株Ish-ikawa,G418筛选稳定并有效表达pRNAT-suv和pRNAT-neg的克隆。用蛋白质印迹法检测稳定表达克隆中Survivin的干扰效果,并用划痕实验、Matrigel体外侵袭实验和蛋白质印迹法评价稳定并有效表达pR-NAT-suv的细胞与阴性对照组细胞运动、侵袭能力以及E-钙黏蛋白表达的差异。结果:稳定表达pRNAT-suv的细胞其Survivin的表达水平低于未转染的细胞和稳定表达pR-NAT-neg的细胞。且稳定表达pRNAT-suv的细胞其运动和侵袭能力均低于未转染的细胞和稳定表达pRNAT-neg的细胞,3组细胞穿透Matrigel基质的细胞数分别为18.60±1.08、50.97±4.87和52.67±4.32,差异有统计学意义,F=63.83,P<0.01。RNA干扰抑制Survivin亦上调了E-钙黏蛋白的表达,差异有统计学意义,F=201.40,P<0.01。结论:RNA干扰抑制Survivin的表达可以降低子宫内膜癌细胞的运动和侵袭能力,可能与上调E-钙黏蛋白的表达有关。Survivin的异常高表达可能促进子宫内膜癌的浸润和转移。更多还原

【Abstract】 OBJECTIVE:To investigate the roles of Survivin on motility and invasion in endometrial cancer cells and its possible mechanisms.METHODS:The recombinant plasmids of pRNAT-suv which contained Survivin siRNA and pRNAT-neg which contained negative control siRNA were used to transfect Ishikawa endometrial cancer cells.The cells were cultured in medium with G418 and the clones stably and effectively expressing pRNAT-suv and pRNAT-neg were selected.Western blot was used to examine the effect of Survivin RNAi.The motility,invasion and E-cadherin expression of cells transfected with pRNAT-suv and pRNAT-neg,or non-transfected cells were observed through wound healing assay,matrigel invasion assay and Western blot.RESULTS:The Survivin expression was lower in cells stably expressing pRNAT-suv than that in non-transfected cells and cells stably expressing pRNAT-neg.The motility and invasion in cells stably expressing pRNAT-suv were also decreased comparing with the non-transfected cells and cells stably expressing pRNAT-neg.The mean numbers of invaded cells in the three groups were 18.60±1.08,50.97±4.87 and 52.67±4.32,respectively(F=63.83,P<0.01).Furthermore,the inhibition of Survivin by RNAi also increased E-cadherin expression(F=201.40,P<0.01).CONCLUSIONS:The inhibition of Survivin can decrease the motility and invasion in Ishikawa cells,which may be associated with the increased E-cadherin expression.The abnormal expression of Survivin can promote the invasion and metastasis in endometrial cancer.更多还原