MicroRNA-23b mediates urokinase and c-met downmodulation and a decreased migration of human hepatocellular carcinoma cells
MicroRNA-23b mediates urokinase and c-met downmodulation and a decreased migration of human hepatocellular carcinoma cells
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DOI：
10.1111/j.1742-4658.2009.07014.x
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发表时间：
2009-06-01
期刊：
影响因子：
5.4
通讯作者：
Barlati, Sergio
中科院分区：
文献类型：
--
作者：
Salvi, Alessandro;Sabelli, Cristiano;Barlati, Sergio
尿激酶型纤溶酶原激活物（uPA）和c-met在肿瘤侵袭和转移中起主要作用。有证据表明，uPA和c-met过表达可能在人肝细胞癌（HCC）中协调。在本研究中，为了了解这些基因的表达是否可能受到人类细胞中特定microRNA（miRs）的共同调控，我们预测智人microRNA-23 b可以识别uPA 3 '-UTR中的两个位点和c-met 3'-UTR中的四个位点。miR-23 b在人肿瘤和正常细胞中的表达分析显示，uPA和c-met表达呈相反趋势，表明uPA和c-met的负调控可能依赖于miR-23 b的表达。转染miR-23 b分子的肝癌细胞（SKHep 1C 3）中靶基因的蛋白表达受到抑制，并导致细胞迁移和增殖能力下降。此外，在人正常AB 2真皮成纤维细胞中转染anti-miR-23 b可上调内源性uPA和c-met的表达。在HCC细胞中，miR-23 b与pGL4.71海肾荧光素酶报告基因构建体（含有推定的uPA和c-met 3 '-UTR靶位点）以及与pGL 3萤火虫荧光素酶表达载体的共转染实验显示相对荧光素酶活性降低。这将表明miR-23 b可以识别uPA和c-met mRNA的3 '-UTR中的靶位点，并在HCC细胞中抑制uPA和c-met的表达。所获得的证据表明，miR-23 b的过表达导致uPA和c-met下调，并降低HCC细胞的迁移和增殖能力。
Urokinase-type plasminogen activator (uPA) and c-met play a major role in cancer invasion and metastasis. Evidence has suggested that uPA and c-met overexpression may be coordinated in human hepatocellular carcinoma (HCC). In the present study, to understand whether the expression of these genes might be coregulated by specific microRNAs (miRs) in human cells, we predicted that Homo sapiens microRNA-23b could recognize two sites in the 3'-UTR of uPA and four sites in the c-met 3'-UTR by the algorithm pictar. The miR-23b expression analysis in human tumor and normal cells revealed an inverse trend with uPA and c-met expression, indicating that uPA and c-met negative regulation might depend on miR-23b expression. Transfection of miR-23b molecules in HCC cells (SKHep1C3) led to inhibition of protein expression of the target genes and caused a decrease in cell migration and proliferation capabilities. Furthermore, anti-miR-23b transfection in human normal AB2 dermal fibroblasts upregulated the expression of endogenous uPA and c-met. Cotransfection experiments in HCC cells of the miR-23b with pGL4.71 Renilla luciferase reporter gene constructs, containing the putative uPA and c-met 3'-UTR target sites, and with the pGL3 firefly luciferase-expressing vector showed a decrease in the relative luciferase activity. This would indicate that miR-23b can recognize target sites in the 3'-UTR of uPA and of c-met mRNAs and translationally repress the expression of uPA and c-met in HCC cells. The evidence obtained shows that overexpression of miR-23b leads to uPA and c-met downregulation and to decreased migration and proliferation abilities of HCC cells.