a Department of Biochemistry and Molecular Biology , Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center , Omaha.
RNA Biol. 2018 Mar 4;15(3):391-403. doi: 10.1080/15476286.2017.1422471. Epub 2018 Feb 1.
MicroRNA dysregulation is a common feature of cancer and due to the promiscuity of microRNA binding this can result in a wide array of genes whose expression is altered. miR-106b is an oncomiR overexpressed in cholangiocarcinoma and its upregulation in this and other cancers often leads to repression of anti-tumorigenic targets. The goal of this study was to identify the miR-106b-regulated gene landscape in cholangiocarcinoma cells using a genome-wide, unbiased mRNA analysis. Through RNA-Seq we found 112 mRNAs significantly repressed by miR-106b. The majority of these genes contain the specific miR-106b seed-binding site. We have validated 11 genes from this set at the mRNA level and demonstrated regulation by miR-106b of 7 proteins. Combined analysis of our miR-106b-regulated mRNA data set plus published reports indicate that miR-106b binding is anchored by G:C pairing in and near the seed. Novel targets Kruppel-like factor 2 (KLF2) and KLF6 were verified both at the mRNA and at the protein level. Further investigation showed regulation of four other KLF family members by miR-106b. We have discovered coordinated repression of multiple members of the KLF family by miR-106b that may play a role in cholangiocarcinoma tumor biology.
miRNA 失调是癌症的一个常见特征，由于 miRNA 结合的随意性，这可能导致广泛的基因表达发生改变。miR-106b 在胆管癌中过表达，其在这种癌症和其他癌症中的上调通常导致抗肿瘤靶基因的抑制。本研究的目的是使用全基因组、无偏 mRNA 分析来鉴定胆管癌细胞中 miR-106b 调节的基因图谱。通过 RNA-Seq，我们发现 112 个 mRNA 被 miR-106b 显著抑制。这些基因中的大多数都包含特定的 miR-106b 种子结合位点。我们已经在 mRNA 水平上验证了这一组中的 11 个基因，并证明了 miR-106b 对 7 种蛋白质的调节。对我们的 miR-106b 调节的 mRNA 数据集加上已发表的报告的综合分析表明，miR-106b 结合是由种子内和附近的 G:C 配对锚定的。新型靶基因 Kruppel 样因子 2 (KLF2) 和 KLF6 在 mRNA 和蛋白质水平上均得到验证。进一步的研究表明，miR-106b 调节了另外四个 KLF 家族成员。我们发现 miR-106b 对 KLF 家族的多个成员进行了协调抑制，这可能在胆管癌肿瘤生物学中发挥作用。
