目的 进一步了解miRNA在膀胱癌中的潜在机制.方法 芯片分析4对人膀胱癌组织和相邻正常组织中的miRNA的表达.并用RT-qPCR来验证两个最上调的miRNA及其靶基因的表达是否符合miRNA/mRNA芯片结果.通过相关性分析和双荧光素酶报告实验推断并验证miR-130b-3p可以靶向PTEN.应用CCK8、EDU、流式细胞术、划痕、Transwell和细胞骨架等实验证明miR-130b可以影响膀胱癌细胞的增殖、凋亡、迁移和侵袭.用Western blot检测PI3K/AKT和整合素β1/FAK信号通路的关键靶蛋白.结果 人膀胱癌中miR-130b-3p表达高于癌旁且与PTEN表达呈负相关.miR-130b3-p可下调PTEN表达,导致PI3K/AKT和整合素β1/FAK信号通路的激活,且与膀胱癌EJ细胞的增殖、迁移和侵袭相关.细胞转染miR-130b-3p抑制剂时,可以重排细胞骨架.结论 本结果揭示miR-130b/PTEN有望用于人膀胱癌诊断和治疗的标志物.%Objective To further understand potential mechanism of miRNAs in bladder cancer .Mtehods Human microarray was used to analyze the expression of miRNAs in four pair human BC cancer tissues and adjacent normal tissues.Then RT-Qpcr was used to verified that the expression of two most up -regulated miRNAs and target genes for results of miRNA/Mrna microarray.Subsequently, it was deduced that miR-130b-3p could target at PTEN through a bioinformatics approach and dual-luciferase reporter assay.CCK8, EDU, flow cytometry, wound healing, Transwell and cytoskeleton assay were applied to prove that miR-130b could affect proliferation, apoptosis, migration and invasion of BC cells.The effects of PTEN regula-ted by miR-130b-3p on the key target protein expression of PI 3K/AKT and integrin β1/FAK signaling pathway were determined by Western blot.Resulst miR-130b-3p was significantly up-regulated and nega-tively correlated with PTEN expression in clinical bladder cancer specimens as compared with normal urothelial tissue.miR-130b-3p mimics could down-regulate PTEN expression, which leads to the activation of PI3K/AKT and integrinβ1/FAK signaling pathway related to cell proliferation , migration and invasion in bladder cancer EJ cells.When the cells were transfected with miR-130b-3p inhibitors, they could be sur- veyed with rearranging cytoskeleton.Conclusions miR-130b/PTEN may be used as a marker for bladder cancer.
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