siRNA 干扰的DKK1低表达对子宫内膜癌Ishikawa细胞侵袭、迁移的影响及机制研究.doc

  siRNA 干扰的 DKK1 低表达对子宫内膜癌 Ishikawa 细胞侵袭、迁移的影响及机制研究 #  5 10 15 20 25 30 35 40 45  伊诺1，华文浩2，付丽华1，周明书1，许仲婷1，李振华1，许艳丽1，易为1，刘敏 1* （1. 首都医科大学附属北京地坛医院妇产科，北京 100015； 2. 首都医科大学附属北京地坛医院检验科，北京 100015） 摘要：目的 探讨 DKK1 siRNA 干扰的 DKK1 低表达对子宫内膜癌 Ishikawa 细胞侵袭、迁移 能力的影响及作用机制。方法 瞬时转染 DKK1 siRNA 至培养的子宫内膜癌 Ishikawa 细胞； 应用实时定量 PCR 和蛋白免疫印迹方法分别检测转染前后细胞中 DKK1 的 mRNA 和蛋白表 达；应用 Transwell 实验检测癌细胞侵袭、迁移能力。应用荧光显微镜观察转染前后 Wnt/ β -catenin 信号通路中活性β -catenin、MMP14 表达情况。结果 DKK1 siRNA 能转染至 Ishikawa 细胞，并使细胞中 DKK1 基因表达下降 21.00 %。DKK1 siRNA 干扰后，细胞中活 性β -catenin、MMP14 表达增高；癌细胞的侵袭能力增强 13.82 %，癌细胞的迁移能力增强 16.39 %。结论 DKK1 具有抑制子宫内膜癌 Ishikawa 细胞侵袭、迁移的作用。Wnt/β -catenin 信号通路参与了 DKK1 这一作用过程。 关键词：肿瘤；DKK1；子宫内膜癌；Ishikawa 细胞系；侵袭；迁移 中图分类号：R737.33 Influence Mechanism of siRNA-mediated DKK1 Low Expression on Invasion and Migration Ability in Endometrial Carcinoma Ishikawa Cell Lines Yi Nuo1, Hua Wenhao2, Fu Lihua1, Zhou Mingshu1, Xu Zhongting1, Li Zhenhua1, Xu Yanli1, Yi Wei1, Liu Min1 (1. Department of Obstetrics and Gynecology, Beijing Ditan Hospital, Capital Medical University, Beijing 100015; 2. Department of Clinical Laboratory, Beijing Ditan Hospital, Capital Medical University, Beijing 100015) Abstract: Objective In this paper the influence mechanism of siRNA-mediated DKK1 low expression on invasion and migration ability was investigated in endometrial carcinoma Ishikawa cell lines. Methods Culture endometrial carcinoma Ishikawa cell lines were transient transfected by DKK1 siRNA; mRNA and protein levels of DKK1 of transfected cells and untransfected cells were revealed by real time PCR and western blot analysis, respectively. The cell invasion and migration ability were detected by Transwell assay. Expression location and profiles of active β-catenin and MMP14 of Wnt/β-catenin signal pathway in cells were detected by immunofluorescence. Results DKK1 siRNA can be transfected into Ishikawa cells and the expression level of DKK1 gene was down-regulated by 21.00 percent. After DKK1 siRNA transfection, the expressi