摘要
目的利用siRNA载体介导的RNA干扰技术靶向沉默β-catenin以阻断Wnt/β-catenin信号通路,观察对子宫内膜异位症裸鼠模型异位子宫内膜血管内皮生长因子(vascular endothelial growth factor,VEGF)和基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)表达的影响,探讨Wnt/β-catenin信号通路促进异位内膜组织粘附、侵袭和转移的作用。方法采用人子宫内膜异体移植方法构建子宫内膜异位症裸鼠模型。将18只子宫内膜异位症裸鼠随机分为3组(n=6):siRNA干扰组、阴性对照组和空白对照组,前两组每日分别腹腔注射100μL siRNA转染复合物(含β-catenin siRNA3μg和转染试剂7.5μg)和阴性对照转染复合物(含阴性对照si RNA3μg和转染试剂7.5μg)1次,共注射5次。末次腹腔注射后24 h断颈处死裸鼠,观察盆、腹腔病灶形成情况,取异位病灶标本。Real time PCR和免疫组织化学检测异位内膜组织β-catenin、VEGF和MMP-9 mRNA和蛋白的表达。结果实验裸鼠共20只,18只成模成活。Real timePCR检测结果显示,与阴性对照组和空白对照组相比,siRNA干扰组β-catenin、VEGF和MMP-9 mRNA表达明显降低(均P<0.05),而后两者之间差异无统计学意义(均P>0.05)。免疫组化结果显示,si RNA干扰组β-catenin、VEGF和MMP-9表达明显减弱,显著低于阴性对照组和空白对照组(均P<0.05),而后两者之间差异无统计学意义(均P>0.05)。结论阻断Wnt/β-catenin信号通路能明显抑制子宫内膜异位症裸鼠模型异位内膜组织VEGF和MMP-9mRNA和蛋白表达。Wnt/β-catenin信号通路可能促进子宫内膜异位症患者在位子宫内膜的异位种植。
Objective To investigate the effect of blockage of Wnt/β-catenin signaling pathway by siRNA interference on the expression of VEGF and MMP-9 in the nude mouse model of endometriosis,and whether the signaling pathway could accelerate the adhesion,invasion and angiogenesis of the implantation of the endometrium of endometriosis.Methods The nude mouse endometriosis model was developed by placing human endometrium fragments into the peritoneal cavity.Eighteen healthy female nude mice were randomly and equally divided into 3 groups（n=6 each）：siRNA group（injecting β-catenin siRNA 3 μg and transfection reagent 7.5 μg）,negative control group（injecting negative fluorescien control siRNA 3 μg and transfection reagent 7.5 μg）and blank control group.Each animal was injected intraperitoneally,daily for 5 days.Twenty-four h after last intraperitoneal injection,animals were sacrificed,and ectopic endometria were collected and homogenized.Real-time PCR and immunohistochemistry were used to detect the expression of β-catenin,VEGF and MMP-9.Results The expression levels of VEGF and MMP-9 mRNA and protein were down-regulated in siRNA group as compared with those in negative control group and blank control group（P0.05）.Conclusion siRNA-mediated inhibition of Wnt/β-catenin limited the expression of VEGF and MMP-9 mRNA and protein in ectopic endometrial of the nude mouse endometriosis model.Wnt/β-catenin signaling pathway may activate implantation of the endometrium of patients with endometriosis.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2011年第1期22-27,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.30750007)
