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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。
Objective: To investigate the regulation of Survivin on the movement and invasion of endometrial cancer cells and its possible mechanism. METHODS: The recombinant plasmid pRNAT-suv containing Survivin-specific siRNA and the negative control plasmid pR-NAT-neg were transfected into endometrial cancer cell line Ish-ikawa respectively. G418 screened stable and effectively expressed pRNAT-suv and pRNAT-neg. clone. Western blotting was used to detect the interference effect of Survivin in stably expressing clones, and to evaluate the cell motility and invasion ability of cells stably and effectively expressing pR-NAT-suv and the negative control group using scratch test, Matrigel in vitro invasion assay and Western blotting. - Differences in cadherin expression. Results: The expression level of Survivin in cells stably expressing pRNAT-suv was lower than that of non-transfected cells and cells stably expressing pR-NAT-neg. The cells that stably expressed pRNAT-suv had lower motor and invasive ability than untransfected cells and cells stably expressing pRNAT-neg. The number of cells that penetrated Matrigel matrix in the three groups was 18.60±1.08 and 50.97±4.87, respectively. 52.67±4.32, the difference was statistically significant, F=63.83, P<0.01. The inhibition of survivin by RNA interference also up-regulated the expression of E-cadherin, and the difference was statistically significant (F=201.40, P<0.01). Conclusion: The inhibition of Survivin expression by RNA interference can reduce the movement and invasion of endometrial cancer cells, which may be related to the up-regulation of E-cadherin expression. Abnormally high expression of Survivin may promote the infiltration and metastasis of endometrial cancer.