miRNA-182-5p promotes human bladder cancer proliferation and migration through the FOXF2/SHH axis
miRNA-182-5p promotes human bladder cancer proliferation and migration through the FOXF2/SHH axis
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miRNA-182-5p通过FOXF2/SHH轴促进人膀胱癌增殖和迁移
DOI：
10.4149/neo_2021_210903n1266
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发表时间：
2022-01-01
期刊：
影响因子：
3
通讯作者：
Li, Yawei
中科院分区：
文献类型：
--
作者：
Zhang, Ze;Wang, Chong;Li, Yawei
越来越多的证据表明，microRNAs （miRNAs）通过与相应mrna的3‘非翻译区（3’-UTRs）结合负向调控基因表达，在膀胱肿瘤的发生发展中起着至关重要的作用。miR-182-5p在膀胱癌（BC）中的作用尚不清楚。因此，本研究旨在阐明miR-182-5p在BC中的功能作用。我们通过三个数据库（TarBase、ENCORI和miRDB）预测miR-182-5p的候选mrna。双荧光素酶报告基因检测和靶标预测证实FOXF2是miR-182-5p的潜在靶标。定量RT-PCR （qRT-PCR）结果显示，内源性miR-182-5p在BC细胞系和BC患者临床样本中表达显著上调。免疫组化（IHC）、western blotting和qRT-PCR检测显示，FOXF2在BC组织和BC细胞系中同时下调表达。功能增益和功能丧失研究表明，过表达miR-182-5p增强了BC细胞的增殖和迁移，而下调miR-182-5p则显示相反的结果。miR-182-5p诱导的作用随着FOXF2表达的恢复而减弱。在BC细胞中，miR-182-5p的上调不仅降低了FOXF2的表达，而且显著增加了Sonic hedgehog （SHH）通路的水平。这些发现表明FOXF2直接结合miR-182-5p， miR-182-5p通过靶向FOXF2在BC发生和转移中发挥肿瘤启动子的作用。此外，miR-182-5p通过下调FOXF2激活SHH通路发挥促癌作用。
Increasing evidence suggests that microRNAs (miRNAs) play critical roles in bladder tumorigenesis and development by combining with the 3' untranslated regions (3'-UTRs) of the corresponding mRNAs to negatively regulate gene expression. The role of miR-182-5p in bladder cancer (BC) remains unclear. Therefore, this study aimed to clarify the functional role of miR-182-5p in BC. We predicted candidate mRNAs for miR-182-5p via three databases (TarBase, ENCORI, and miRDB). Dual-luciferase reporter assays and target prediction confirmed FOXF2 as a potential target of miR-182-5p. Quantitative RT-PCR (qRT-PCR) showed that endogenous miR-182-5p expression was significantly upregulated in BC cell lines and clinical samples of BC patients. IHC, western blotting, and qRT-PCR assays indicated that FOXF2 expression was concurrently downregulated in BC tissues and BC cell lines. Gain- and loss-of-function studies showed that the overexpression of miR-182-5p enhanced the proliferation and migration of BC cells, while the downregulation of miR-182-5p showed the opposite results. The effects induced by miR-182-5p were attenuated with the restoration of FOXF2 expression. In BC cells, the upregulation of miR-182-5p not only decreased FOXF2 expression but also markedly increased Sonic hedgehog (SHH) pathway levels. These findings suggested that FOXF2 directly binds to miR-182-5p and that miR-182-5p acts as a tumor promoter in BC genesis and metastasis by targeting FOXF2. In addition, miR-182-5p plays a pro-cancer role by downregulating FOXF2 and activating the SHH pathway.