We established a semi-high-throughput screening platform using hyperactive (hyPB) transposons (designated as ) to identify microRNAs (s) that inhibit hepatocellular carcinoma (HCC) development , following overexpression in hepatocytes. s encoding six different s from the cluster and nine s from outside this cluster were transfected into mouse livers that were chemically induced to develop HCC. In this slow-onset HCC model, significantly inhibited HCC. Next, we developed a more aggressive HCC model by overexpression of oncogenic Harvey rat sarcoma viral oncogene homolog ) and oncogenes that accelerated HCC development after only 6 weeks. The tumor suppressor effect of could be demonstrated even in this rapid-onset / HCC model, consistent with significantly prolonged survival and decreased HCC tumor burden. Comprehensive RNA expression profiling of 95 selected genes typically associated with HCC development revealed differentially expressed genes and functional pathways that were associated with -mediated HCC suppression. To our knowledge, this is the first study establishing a direct causal relationship between overexpression and liver cancer inhibition . Moreover, these results demonstrate that hepatocyte-specific hyPB transposons are an efficient platform to screen and identify s that affect overall survival and HCC tumor regression.

中文翻译：

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使用肝细胞特异性高活性piggyBac转座子验证miR-20a作为肝癌中的肿瘤抑制基因


我们建立了一个半高通量筛选平台，使用高活性（hyPB）转座子（指定为 ）来鉴定在肝细胞中过度表达后抑制肝细胞癌（HCC）发展的 microRNA。编码该簇中 6 个不同 s 和该簇外 9 个不同 s 的 s 被转染到小鼠肝脏中，并通过化学方法诱导小鼠肝脏发生 HCC。在这种缓慢发作的 HCC 模型中，显着抑制 HCC。接下来，我们通过过度表达致癌的哈维大鼠肉瘤病毒癌基因同源物和仅 6 周后加速 HCC 发展的癌基因，开发了更具侵袭性的 HCC 模型。即使在这种快速起病的 HCC 模型中也能证明其抑癌作用，这与显着延长生存期和降低 HCC 肿瘤负荷相一致。对通常与 HCC 发展相关的 95 个选定基因的综合 RNA 表达谱揭示了与介导的 HCC 抑制相关的差异表达基因和功能途径。据我们所知，这是第一项建立过度表达与肝癌抑制之间直接因果关系的研究。此外，这些结果表明，肝细胞特异性 hyPB 转座子是筛选和识别影响总体生存和 HCC 肿瘤消退的有效平台。