Wang Yanmei, Sun Guiying, Wang Chenghong, Guo Wenchen, Tang Qianqing, Wang Meng
Department of Urology, Jining First People's Hospital, Jining, Shandong Province, 272000, P.R. China.
J BUON. 2018 Sep-Oct;23(5):1492-1499.
MicroRNA (miR)-194-5p is downregulated in bladder cancer (BC), but its role in BC has not been determined mechanistically.
The expression levels of miR-194-5p and E2F transcription factor 3 (E2F3) were determined by means of quantitative reverse transcription and polymerase chain reaction in BC specimens. In addition, T24 BC cells were transfected with a miR-194-5p mimic, a miR-194-5p inhibitor, or E2F3 small interfering (si)RNA, and the level of E2F3 protein expressed by these cells was assessed by western blotting. A dual luciferase reporter assay was applied to verify the binding site between miR-194-5p and the 3' untranslated region of E2F3. Transwell assays were performed to examine cell migration and invasion.
Dysregulation of miR-194-5p in BC was closely associated with node metastasis and differentiation. In BC specimens and cell lines, miR-194-5p mRNA was downregulated, while E2F3 mRNA was upregulated. Overexpression of miR-194-5p suppressed the expression of E2F3 mRNA and protein. By regulating E2F3, miR-194-5p inhibited cell migration and invasion in BC. Treatment of BC cells with E2F3 siRNA had the same effect as did overexpression of miR-194-5p.
MiR-194-5p directly targets E2F3 and inhibits cell migration and invasion in BC.
微小RNA（miR）-194-5p在膀胱癌（BC）中表达下调，但其在BC中的作用尚未得到机制上的确定。
通过定量逆转录和聚合酶链反应测定BC标本中miR-194-5p和E2F转录因子3（E2F3）的表达水平。此外，用miR-194-5p模拟物、miR-194-5p抑制剂或E2F3小干扰（si）RNA转染T24 BC细胞，并通过蛋白质印迹法评估这些细胞表达的E2F3蛋白水平。应用双荧光素酶报告基因测定法验证miR-194-5p与E2F3的3'非翻译区之间的结合位点。进行Transwell测定以检查细胞迁移和侵袭。
BC中miR-194-5p的失调与淋巴结转移和分化密切相关。在BC标本和细胞系中，miR-194-5p mRNA下调，而E2F3 mRNA上调。miR-194-5p的过表达抑制了E2F3 mRNA和蛋白的表达。通过调节E2F3，miR-194-5p抑制了BC中的细胞迁移和侵袭。用E2F3 siRNA处理BC细胞与miR-194-5p过表达具有相同的效果。
MiR-194-5p直接靶向E2F3并抑制BC中的细胞迁移和侵袭。
