中文摘要：

miR-130家族在癌的进展中发挥作用;然而,其家族成员在膀胱癌中的作用尚罕见报告。本研究证明,抑制miR-130家族成员miR-130b-3p表达促进PTEN表达,诱导细胞凋亡,抑制膀胱癌细胞增殖。首先,我们采用微阵列对来自膀胱癌患者的4对膀胱癌和癌旁组织进行了miRNA和mRNA组学分析,发现miR-130b-3p和miR-106b-3p在膀胱癌组织高表达,而miR-99a-3p、miR-199a-5p和miR-145-3p低表达。RT-q PCR检测30对膀胱癌组织5种miRNA的表达与微阵列中的表达状况一致。此外,我们在mRNA组学分析中还发现,miR-130b-3p在膀胱癌组织的高表达与PTEN表达呈负相关。为此,在后续研究中集中探索了miR-130b-3p在膀胱癌中的作用及其作用机制。生物信息学及荧光素酶报告结果证明,miR-130b-3p可直接结合PTEN的3＇-UTR,靶向抑制PTEN的表达。转染结合CCK-8、EDU、流式分析、划痕及Transwell小室实验显示,转染miR-130b-3p模拟物可明显促进膀胱癌T24细胞的增殖、迁移及侵袭能力;相反,转染miR-130b-3p抑制物可明显诱导T24细胞凋亡。鬼笔环肽染色揭示,转染miR-130b-3p模拟物可促进细胞骨架形成,而转染miR-130b-3p抑制物抑制细胞骨架形成。Western印迹证明,转染miR-130b-3p可下调PTEN在T24细胞的表达,上调p-PI3K、p-AKT、p-FAK和整合素β1的表达;而转染miR-130b-3p抑制物上调PTEN的表达,下调p-PI3K、p-AKT、p-FAK和整合素β1的表达。上述结果提示,miR-130b-3p通过抑制PTEN表达,激活PI3K-AKT及整合素β1/FAK信号通路,在膀胱癌中发挥癌基因样作用;相反,抑制miR-130b-3p可上调PTEN表达,抑制PI3K-AKT及整合素β1/FAK信号通路的激活,诱导凋亡,抑制膀胱癌细胞的增殖、迁移和侵袭能力。我们的结果还提示,miR-130b-3p作为可能的临床标志物,对膀胱癌的诊断、靶向治疗具有潜在的应用价值。

英文摘要：

The miR-130 family plays a role in the progression of cancer; however, the function of its family members in bladder cancer is still rarely reported. This study demonstrates that down-regulation of miR-130b-3p promotes the expression of PTEN, induces apoptosis and inhibits proliferation of bladder cancer cells. Firstly, we found miR-130b-3p and miR-106b-3p were highly expressed in bladder cancer tissues compared with the adjacent tissues from four bladder cancer patients with grade 2 （G2） T2 using microarray analysis, while the expression of miR-99a-3p, miR-199a-5p and miR-145-3p were downregulated. RT-qPCR assay showed that the levels of these 5 miRNAs in 30 eases of bladder cancer were eonsistent with the data of mieroarray analysis. We also found that miR-130b-3p expression was negatively correlated with PTEN expression in bladder cancer tissues. Therefore, we focused on the roles and mechanisms of miR-130b-3p in bladder cancer. Bioinformatics and luciferase reporter assay showed that miR-130b-3p could directly bind to the 3＇-UTR of PTEN and inhibited PTEN expression. CCK-8, EDU, flow cytometry, wound healing and transwell assays showed that transfection with miR-130b-3p mimics eould significantly promote the proliferation, migration and invasion of bladder cancer T24 cells; miR-130b-3p inhibitors could induce apoptosis of T24 cells. Phalloidin staining revealed that transfeetion with miR-130b-3p mimics promoted eytoskeletal formation, while transfection with miR-130b-3p inhibitors suppressed stress fiber formation. Results showed that miR-130b-3p could down-regulate the expression of PTEN in T24 cells and up-regulate the expression of p-PI3K, p-AKT, p-FAK and integrin [31 by Western blotting; while miR-130b-3p inhibitors could up-regulate the expression of PTEN and down-regulate the expression of p-PI3K, p-AKT, p-FAK and integrin [31. In contrast, miR-130b-3p inhibited the PI3K-AKT and integrin β1/FAK signaling pathways by up-regulating the expression of PTEN, thus leading to promotion of apoptosis and