Involvement of aberrantly activated HOTAIR/EZH2/miR-193a feedback loop in progression of prostate cancer.
Involvement of aberrantly activated HOTAIR/EZH2/miR-193a feedback loop in progression of prostate cancer.
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异常激活的 HOTAIR/EZH2/miR-193a 反馈环路参与前列腺癌的进展
DOI：
10.1186/s13046-017-0629-7
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发表时间：
2017-11-15
期刊：
影响因子：
--
通讯作者：
Chen M
中科院分区：
文献类型：
--
作者：
Ling Z;Wang X;Tao T;Zhang L;Guan H;You Z;Lu K;Zhang G;Chen S;Wu J;Qian J;Liu H;Xu B;Chen M
背景虽然雄激素剥夺治疗是前列腺癌（prostate cancer，PCa）的标准治疗方法，但大多数患者不可避免地发展为去势抵抗性前列腺癌（castra-resistant prostate cancer，CRPC），这是导致PCa死亡的主要原因。因此，鉴定调节癌症进展的新分子机制和实现靶向治疗的新见解对于改善PCa患者的益处是必要的。本研究旨在探讨HOTAIR/EZH 2/miR-193 a反馈环在前列腺癌进展中的作用及调控机制。采用CCK-8法、集落形成法、侵袭法、迁移法、流式细胞术、异种移植模型和基因集富集分析法检测和分析miR-193 a的生物学功能。然后，我们通过质粒、慢病毒和小干扰RNA（siRNA）评估HOTAIR和EZH 2在调节miR-193 a表达中的作用。进行荧光素酶报告基因测定和染色质免疫沉淀测定以检测EZH 2和HOTAIR对miR-193 a的转录激活。此外，进行qRT-PCR和荧光素酶报告基因测定以检查miR-193 a控制PCa中HOTAIR表达的调节作用。最后，采用原位杂交和免疫组化方法分析HOTAIR、EZH 2和miR-193 a表达的相关性。体外研究表明，miR-193 a抑制PCa细胞生长，抑制迁移和侵袭，并促进凋亡;体内结果表明，慢病毒介导的miR-193 a过表达显著降低PCa异种移植肿瘤生长。重要的是，我们发现EZH 2与HOTAIR偶联通过在PC 3和DU 145细胞中miR-193 a启动子处的H3 K27的三甲基化来抑制miR-193 a表达。有趣的是，进一步的证据表明，miR-193 a直接靶向HOTAIR，显示在miR-193 a过表达的细胞和组织中HOTAIR水平显著降低。结论miR-193 a在CRPC中具有抑癌作用，HOTAIR/EZH 2/miR-193 a的自调节反馈环在PCa的发生发展中起重要作用。针对这种异常激活的反馈回路可能提供一种潜在的治疗策略。
BackgroundThough androgen deprivation therapy is the standard treatment for prostate cancer (PCa), most patients would inevitably progress to castration-resistant prostate cancer (CRPC) which is the main cause of PCa death. Therefore, the identification of novel molecular mechanism regulating cancer progression and achievement of new insight into target therapy would be necessary for improving the benefits of PCa patients. This study aims to study the function and regulatory mechanism of HOTAIR/EZH2/miR-193a feedback loop in PCa progression.MethodsMSKCC and TCGA datasets were used to identify miR-193a expression profile in PCa. Cell Counting Kit-8 (CCK-8) assays, colony formation, invasion, migration, flow cytometry, a xenograft model and Gene Set Enrichment Analysis were used to detect and analyze the biological function of miR-193a. Then, we assessed the role of HOTAIR and EZH2 in regulation of miR-193a expression by using plasmid, lentivirus and small interfering RNA (siRNA). Luciferase reporter assays and chromatin immunoprecipitation assays were performed to detect the transcriptional activation of miR-193a by EZH2 and HOTAIR. Further, qRT-PCR and luciferase reporter assays were conducted to examine the regulatory role of miR-193a controlling the HOTAIR expression in PCa. Finally, the correlation between HOTAIR, EZH2 and miR-193a expression were analyzed using In situ hybridization and immunohistochemistry.ResultsWe found that miR-193a was significantly downregulated in metastatic PCa through mining MSKCC and TCGA datasets. In vitro studies revealed that miR-193a inhibited PCa cell growth, suppressed migration and invasion, and promoted apoptosis; in vivo results demonstrated that overexpression of miR-193a mediated by lentivirus dramatically reduced PCa xenograft tumor growth. Importantly, we found EZH2 coupled with HOTAIR to repress miR-193a expression through trimethylation of H3K27 at miR-193a promoter in PC3 and DU145 cells. Interestingly, further evidence illustrated that miR-193a directly targets HOTAIR showing as significantly reduced HOTAIR level in miR-193a overexpressed cells and tissues. The expression level of miR-193a was inversely associated with that of HOTAIR and EZH2 in PCa.ConclusionThis study firstly demonstrated that miR-193a acted as tumor suppressor in CRPC and the autoregulatory feedback loop of HOTAIR/EZH2/miR-193a served an important mechanism in PCa development. Targeting this aberrantly activated feedback loop may provide a potential therapeutic strategy.
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影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1016/j.bbcan.2015.07.001
发表时间：
2015-08
期刊：
Biochimica et biophysica acta
影响因子：
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影响因子：
11.2
作者：
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