microRNA-145 promotes differentiation in human urothelial carcinoma through down-regulation of syndecan-1.
microRNA-145 promotes differentiation in human urothelial carcinoma through down-regulation of syndecan-1.
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DOI：
10.1186/s12885-015-1846-0
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发表时间：
2015-10-29
期刊：
影响因子：
3.8
通讯作者：
Konishi N
中科院分区：
文献类型：
--
作者：
Fujii T;Shimada K;Tatsumi Y;Hatakeyama K;Obayashi C;Fujimoto K;Konishi N
需要一种新的膀胱癌分子标记物作为诊断工具或治疗靶点。潜在的标记物包括 microRNA (miRNA)，它们是长度为 19-24 nt 的短、低分子量 RNA，可调节与各种癌症中的细胞增殖、分化和发育相关的基因。在这项研究中，我们研究了 miR-145 促进尿路上皮癌细胞存活和分化为多个谱系的分子机制。我们发现 miR-145 可以调节 syndecan-1（一种硫酸肝素蛋白聚糖）的表达。通过 MTS 测定评估人尿路上皮癌细胞系 T24 和 KU7 中的细胞增殖。分别通过衰老相关β-半乳糖苷酶（SA-β-gal）和TUNEL测定来测量细胞衰老和凋亡。定量 RT-PCR 用于测量各种基因的 mRNA 表达，包括 Syndecan-1、干细胞因子以及分化为鳞状细胞、腺细胞或神经内分泌细胞的标记物。 miR-145 的过表达诱导细胞衰老，从而显着抑制 T24 和 KU7 细胞的细胞增殖。 Syndecan-1 表达减少，而 SOX2、NANOG、OCT4 和 E2F3 等干细胞标记物增加。 miR-145 还上调分化为鳞状细胞（p63、TP63 和 CK5）、腺细胞（MUC-1、MUC-2 和 MUC-5 AC）和神经内分泌细胞（NSE 和 UCHL-1）的标志物。最后，miR-145 的表达在高级别尿路上皮癌中下调，但在低级别肿瘤中则不然。结果表明，miR-145 抑制 syndecan-1，并通过这种机制上调干细胞因子并诱导细胞衰老和分化。我们认为 miR-145 可能赋予尿路上皮癌细胞类似干细胞的特性，从而促进其分化为多种细胞类型。本文的在线版本 (doi:10.1186/s12885-015-1846-0) 包含补充材料，可供授权用户使用。
A new molecular marker of carcinoma in the urinary bladder is needed as a diagnostic tool or as a therapeutic target. Potential markers include microRNAs (miRNAs), which are short, low molecular weight RNAs 19–24 nt long that regulate genes associated with cell proliferation, differentiation, and development in various cancers. In this study, we investigated the molecular mechanisms by which miR-145 promotes survival of urothelial carcinoma cells and differentiation into multiple lineages. We found miR-145 to regulate expression of syndecan-1, a heparin sulfate proteoglycan. Cell proliferation in the human urothelial carcinoma cell lines T24 and KU7 was assessed by MTS assay. Cellular senescence and apoptosis were measured by senescence-associated β-galactosidase (SA-β-gal) and TUNEL assay, respectively. Quantitative RT-PCR was used to measure mRNA expression of various genes, including syndecan-1, stem cell factors, and markers of differentiation into squamous, glandular, or neuroendocrine cells. Overexpression of miR-145 induced cell senescence, and thus significantly inhibited cell proliferation in T24 and KU7 cells. Syndecan-1 expression diminished, whereas stem cell markers such as SOX2, NANOG, OCT4, and E2F3 increased. miR-145 also up-regulated markers of differentiation into squamous (p63, TP63, and CK5), glandular (MUC-1, MUC-2, and MUC-5 AC), and neuroendocrine cells (NSE and UCHL-1). Finally, expression of miR-145 was down-regulated in high-grade urothelial carcinomas, but not in low-grade tumors. Results indicate that miR-145 suppresses syndecan-1 and, by this mechanism, up-regulates stem cell factors and induces cell senescence and differentiation. We propose that miR-145 may confer stem cell-like properties on urothelial carcinoma cells and thus facilitate differentiation into multiple cell types. The online version of this article (doi:10.1186/s12885-015-1846-0) contains supplementary material, which is available to authorized users.