【作者】 刘儒彪；

【导师】 刘义；

【作者基本信息】 华中科技大学 ， 妇产科， 2010， 硕士

【摘要】 目的:通过观察人子宫内膜异位症在位子宫内膜间质细胞(endometrial stroma cell, ESC)β-catenin的表达及利用siRNA(small interfering RNA, siRNA)靶向沉默β-catenin基因以阻断Wnt/β-catenin信号通路来观察靶基因血管内皮生长因子(vascularendothelial growth factor, VEGF)和基质金属蛋白酶-9(matrix metalloproteinase-9, MMP-9)表达的变化,探讨Wnt/β-catenin信号通路在子宫内膜异位症发生机制中的作用。方法:采用随机、对照设计,将体外分离、培养的子宫内膜异位症在位内膜间质细胞24例分成3组(n=8):空白组、阴性荧光组(以含阴性荧光siRNA 100pmol和脂质体lip2000 5μl的混合物转染)以及siRNA干扰组(以含β-catenin siRNA 100pmol和脂质体lip2000 5μl的混合物转染),并以非子宫内膜异位症子宫内膜间质细胞10例作为正常对照组。利用Real time RT-PCR和western blot方法分别检测子宫内膜异位症在位内膜空白组和非子宫内膜异位症正常对照组β-catenin的表达。转染24h后,用以上方法分别检测子宫内膜异位症空白组、阴性荧光组和siRNA干扰组β-catenin、VEGF以及MMP-9的表达。结果:子宫内膜异位症空白组β-catenin mRNA和蛋白的表达明显高于非子宫内膜异位症正常对照组(P<0.05)。子宫内膜异位症siRNA干扰组β-catenin、VEGF以及MMP-9 mRNA和蛋白的表达明显低于空白组和阴性荧光组(P<0.05),而空白组与阴性荧光组之间无显著差异(P>0.05)。结论:子宫内膜异位症在位内膜间质细胞β-catenin的表达明显高于非子宫内膜异位症患者,阻断Wnt/β-catenin信号通路后,其靶基因VEGF、MMP-9的表达明显受到抑制。因此,Wnt/β-catenin信号通路可能在子宫内膜异位症的发生机制中起重要作用。更多还原

【Abstract】 Objective: Endometriosis is a polygenic disease with a complex and multifactorial aetiology and a common and frequently encountered disease of women of reproductive age. We tested the hypothesis that Wnt/β-catenin signal pathway plays a role on pathogenesis of endometriosis through detecting expression ofβ-catenin in endometrial stromal cells from eutopic endometrium of women with endometriosis and down-stream target gene VEGF, MMP-9 after silencingβ-catenin gene.Methods: Endometrial stromal cells from eutopic endometrium of 24 women with endometrisis(diagnosed by pathology eventually) were divided into 3 subgroups at random: blank subgroup(n=8), negative subgroup(transfection by negative fluorescien control siRNA, n=8) and siRNA interference subgroup(transfection byβ-catenin siRNA, n=8). Then levels ofβ-catenin, VEGF and MMP-9 were determined by Real time RT-PCR and western blot. Endometrial stromal cells from 10 women without endometriosis were as controls.Results: Blank subgroup were found to have higher levels ofβ-catenin than controls(P﹤0.05). In a subgroup analysis of blank, negative and siRNA subgroup, significant differences were found in levels ofβ-catenin, VEGF and MMP-9. Levels ofβ-catenin, VEGF and MMP-9 were significantly lower in siRNA interference subgroup(P﹤0.05), however, there were no differences between blank and negative subgroup(P>0.05).Conclusion:β-catenin expression in endometrial stromal cells from eutopic endometrium of women with endometriosis is increased significantly compared with women withou endometriosis. After blockage of Wnt/β-catenin signal pathway, expression of down-stream target gene VEGF and MMP-9 are suppressed dramaticlly. So, the results of the present study suggest that Wnt/β-catenin signal pathway plays an important role on pathogenesis of endometriosis.更多还原