Department of General Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, China.
Dig Dis Sci. 2012 May;57(5):1171-80. doi: 10.1007/s10620-012-2099-2. Epub 2012 Feb 29.
MicroRNAs (miRNA) are a group of noncoding small RNAs that repress mRNA expression or induce mRNA degradation by binding to the 3'-untranslated regions of mRNAs. MiRNAs have been connected closely with the development of cancers such as hepatocellular carcinoma (HCC). However, the overexpression of microRNA-301a (miR-301a) has seldom been connected with tumorigenesis in HCC.
This study aims to characterize the function of upregulated miR-301a in HCC and show how the downstream growth arrest-specific homeobox (Gax) is negatively regulated by miR-301a.
The expression of miR-301a and Gax was detected using real-time PCR on HCC tissues and adjacent non-tumorous tissues. The luciferase reporter assay was used to assess Gax as a target of miR-301a. The nuclear factor κB (NF-κB) was measured by western blot after inhibiting miR-301a and enhancing Gax. The functions of miR-301a in vivo in HCC cells were measured by migration and invasion assays and flow cytometry.
MiR-301a was significantly upregulated and Gax was downregulated in HCC samples compared with in the matching nontumoral tissues. Inhibiting miR-301a expression caused the upregulation of Gax and repressed NF-κB expression. We have shown that miR-301a plays an important role in increasing proliferation, migration and invasion and in inhibiting apoptosis of HCC cells.
miR-301a is frequently upregulated in HCC and modulates NF-κB expression by negatively regulating Gax.
MicroRNAs（miRNA）是一组非编码的小 RNA，通过与 mRNA 的 3'非翻译区结合来抑制 mRNA 的表达或诱导 mRNA 的降解。miRNA 与肝癌（HCC）等癌症的发展密切相关。然而，miRNA-301a 的过表达与 HCC 中的肿瘤发生很少有关。
本研究旨在表征上调的 miR-301a 在 HCC 中的功能，并展示下游生长停滞特异性同源盒（Gax）如何被 miR-301a 负调控。
使用实时 PCR 检测 HCC 组织和相邻非肿瘤组织中 miR-301a 和 Gax 的表达。荧光素酶报告基因检测用于评估 Gax 作为 miR-301a 的靶标。抑制 miR-301a 和增强 Gax 后，通过 Western blot 测量核因子 kappa B（NF-κB）。通过迁移和侵袭测定以及流式细胞术测量 miR-301a 在 HCC 细胞体内的功能。
与匹配的非肿瘤组织相比，miR-301a 在 HCC 样本中显著上调，而 Gax 下调。抑制 miR-301a 的表达导致 Gax 的上调并抑制 NF-κB 的表达。我们已经表明，miR-301a 在增加 HCC 细胞的增殖、迁移和侵袭以及抑制细胞凋亡方面发挥重要作用。
miR-301a 在 HCC 中频繁上调，并通过负调控 Gax 来调节 NF-κB 的表达。
