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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。
The miR-130 family plays a role in cancer progression; however, the role of its family members in bladder cancer is rarely reported. This study demonstrated that inhibition of miR-130b-3p expression in miR-130 family members promotes PTEN expression, induces apoptosis and inhibits bladder cancer cell proliferation. First, we performed miRNA and mRNA analysis of four pairs of bladder and para-cancerous tissues from bladder cancer patients using microarrays and found that miR-130b-3p and miR-106b-3p were highly expressed in bladder cancer tissues, whereas miR -99a-3p, miR-199a-5p and miR-145-3p. RT-q PCR detected 30 pairs of bladder cancer tissue five miRNA expression and microarray expression status. In addition, we also found in mRNA analysis that the high expression of miR-130b-3p in bladder cancer tissues was negatively correlated with the expression of PTEN. To this end, in the follow-up study focused on exploring miR-130b-3p in bladder cancer and its mechanism of action. Bioinformatics and luciferase reporter results demonstrate that miR-130b-3p binds directly to the 3’-UTR of PTEN and targets the inhibition of PTEN expression. Transfection combined with CCK-8, EDU, flow cytometry, scratch and Transwell chamber experiments showed that transfection of miR-130b-3p mimics significantly promoted the proliferation, migration and invasion of bladder cancer T24 cells; -130b-3p inhibitor can significantly induce T24 cell apoptosis. Phalloidin staining revealed that transfection of miR-130b-3p mimics promoted cytoskeletal formation, whereas miR-130b-3p transfected miR-130b-3p inhibited cytoskeletal formation. Western blotting showed that miR-130b-3p transfected miR-130b-3p down-regulated the expression of PTEN in T24 cells and upregulated the expression of p-PI3K, p-AKT, p-FAK and integrinβ1; PTEN expression, down-regulation of p-PI3K, p-AKT, p-FAK and integrin β1 expression. The above results suggest that miR-130b-3p can play an oncogene-like role in bladder cancer by inhibiting PTEN expression and activating PI3K-AKT and integrinβ1 / FAK signaling. In contrast, miR-130b-3p up-regulates PTEN expression by inhibiting miR- Inhibits the activation of PI3K-AKT and integrin β1 / FAK signaling pathway, induces apoptosis and inhibits the proliferation, migration and invasion of bladder cancer cells. Our results also suggest that miR-130b-3p has potential value as a potential clinical marker for diagnosis and targeted therapy of bladder cancer.