Effect of microRNA-200a on epithelial-mesenchymal transition of bladder cancer cells through targeting β-catenin
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摘要: 目的:探讨miR-200a靶向β-catenin对膀胱癌细胞上皮-间质转换(EMT)的影响。方法:TOP/FOP闪光荧光素酶法鉴定miR-200a对β-catenin信号依赖性;使用荧光素酶法测定miR-200a对β-catenin编码基因CTNNB1的3'非翻译区(3'-UTR)的靶向作用;划痕试验和Transwell检测miR-200a对膀胱癌细胞5637的细胞活力、转移和侵袭影响。Western blotting确定miR-200a对Wnt/β-catenin信号下游分子的效应。结果:miR-200a可直接与β-catenin编码基因CTNNB1的3'UTR相互作用,并抑制β-catenin的表达。miR-200a抑制了5637细胞的活力、转移和侵袭。结论:miR-200a通过靶向β-catenin抑制了膀胱癌细胞5637的生物学功能。
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关键词:
- 膀胱癌 /
- 上皮-间质转换 /
- miR-200a /
- Wnt/β-catenin信号通路
Abstract: Objective: To determine the effect of miR-200 a on the epithelial to mesenchymal transition(EMT) within bladder cancer cells. Method: TOP/FOP flash luciferase assays were employed to identify the effect of miR-200 a on the β-catenin activity. A 3' untranslated region(3'-UTR) luciferase assay was used to determine the target genes of miR-200 a. Assays of cell viability, invasion and wound healing were enabled functional analyses after miR-200 a transfection. Western blotting was employed to determine downstream effects of mine proteins post miR-200 a transfection. Result: Our data showed that miR-200 a can directly interact with the 3'-UTR of β-catenin encoding gene CTNNB1 to suppress β-catenin expression. MiR-200 a also suppressed the biological behaviors of 5637 cells, including cell viability, invasion and wound healing.Conclusion: MiR-200 a suppresses biological function of bladder cancer 5637 cells via targeting-catenin. -
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