Upregulation of microRNA-96 and its oncogenic functions by targeting CDKN1A in bladder cancer.
Upregulation of microRNA-96 and its oncogenic functions by targeting CDKN1A in bladder cancer.
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DOI：
10.1186/s12935-015-0235-8
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发表时间：
2015
影响因子：
5.8
通讯作者：
Gu S
中科院分区：
文献类型：
--
作者：
Wu Z;Liu K;Wang Y;Xu Z;Meng J;Gu S
与正常膀胱组织相比，全基因组miRNA表达谱已将microRNA（miR）-96鉴定为临床膀胱癌（BC）组织中上调的miRNA之一。本研究的目的是确认miR-96在BC组织中的表达模式，并研究其在癌发生中的作用。采用实时荧光定量PCR方法检测60例乳腺癌组织和40例正常对照组织中miR-96的表达水平。采用生物信息学预测结合荧光素酶报告基因分析验证细胞周期蛋白依赖性激酶抑制剂CDKN1A是否为miR-96的潜在靶基因。采用细胞计数试剂盒-8和细胞凋亡实验检测miR-96-CDKN1A轴对BC细胞增殖和凋亡的影响。我们验证了miR-96在人BC组织和细胞系中均显著增加。根据miRTarBase数据，CDKN1A可能是miR-96的候选靶基因。另外，荧光素酶报告基因和Western blot实验分别证实miR-96可与CDKN1A mRNA 3 ′ UTR的种子区结合，并显著降低CDKN1A蛋白的表达水平。此外，我们发现抑制miR-96的表达显著降低了BC细胞系的细胞增殖并促进了细胞凋亡，这与引入不含3 ′ UTR的CDKN1A cDNA恢复miR-96后观察到的结果一致。我们的数据显示，miR-96可能在BC中作为致癌miRNA发挥作用。miR-96的上调可能部分通过抑制BC细胞中CDKN1A蛋白的表达而导致侵袭性恶性肿瘤。
Genome-wide miRNA expression profile has identified microRNA (miR)-96 as one of upregulated miRNAs in clinical bladder cancer (BC) tissues compared to normal bladder tissues. The aim of this study was to confirm the expression pattern of miR-96 in BC tissues and to investigate its involvement in carcinogenesis. Quantitative real-time PCR was performed to detect the expression levels of miR-96 in 60 BC and 40 normal control tissues. Bioinformatics prediction combined with luciferase reporter assay were used to verify whether the cyclin-dependent kinase inhibitor CDKN1A was a potential target gene of miR-96. Cell counting kit-8 and apoptosis assays were further performed to evaluate the effects of miR-96-CDKN1A axis on cell proliferation and apoptosis of BC cell lines. We validated that miR-96 was significantly increased in both human BC tissues and cell lines. According to the data of miRTarBase, CDKN1A might be a candidate target gene of miR-96. In addition, luciferase reporter and Western blot assays respectively demonstrated that miR-96 could bind to the putative seed region in CDKN1A mRNA 3′UTR, and significantly reduce the expression level of CDKN1A protein. Moreover, we found that the inhibition of miR-96 expression remarkably decreased cell proliferation and promoted cell apoptosis of BC cell lines, which was consistent with the findings observed following the introduction of CDKN1A cDNA without 3′UTR restored miR-96. Our data reveal that miR-96 may function as an onco-miRNA in BC. Upregulation of miR-96 may contribute to aggressive malignancy partly through suppressing CDKN1A protein expression in BC cells.