Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, PR China.
Department of General Surgery, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, PR China.
Biochem Biophys Res Commun. 2018 Nov 17;506(1):189-193. doi: 10.1016/j.bbrc.2018.10.038. Epub 2018 Oct 19.
LncRNA has been shown to associates with the initiation and progression of hepatocellular carcinoma (HCC). Recently, some studies showed that HANR function as an oncogene in HCC; however, the detailed mechanism of HANR-regulated HCC tumorigenesis and progression needs to be elucidated.
We used RT-qPCR method to probe genes expression. MTT assay, wound healing assay and transwell invasion assay were utilized to examine proliferation and migration and invasion abilities of HepG2 cells. Xenograft tumor experiment was used to show the growth of tumors in vivo.
HANR was evidently upregulated in HCC tumors and cells compared to normal tissues and cells. Besides, HANR knockdown induces attenuated cell proliferation, migration, invasion of HCC cells. By bioinformatic analysis and dual luciferase reporter assay, we found that miR-214 was the downstream target of HANR. Furthermore, miR-214 inhibitor largely enhanced tumor phenotypes of HCC cells regulated by HANR knockdown. HANR and miR-214 regulated the EZH2, then affecting TGFBR2 level. Finally, we demonstrated that EZH2 overexpression could greatly rescue HANR knockdown or miR-214 mimic-induced HCC tumorigenesis and progression.
In this study, we report a newly identified regulatory mechanism HANR/miR-214/EZH2/TGF-β axis, which is implicated in tumorigenesis and progression of HCC. Our findings suggest that HANR facilitates the development of therapeutical strategies or diagnostic markers by targeting HANR.
长链非编码 RNA（lncRNA）已被证明与肝细胞癌（HCC）的发生和发展有关。最近的一些研究表明，HANR 在 HCC 中作为癌基因发挥作用；然而，HANR 调节 HCC 肿瘤发生和进展的详细机制仍需阐明。
我们使用 RT-qPCR 方法来探测基因表达。MTT 测定、划痕愈合测定和 Transwell 侵袭测定用于检测 HepG2 细胞的增殖、迁移和侵袭能力。异种移植肿瘤实验用于显示体内肿瘤的生长。
与正常组织和细胞相比，HANR 在 HCC 肿瘤和细胞中明显上调。此外，HANR 敲低可诱导 HCC 细胞增殖、迁移和侵袭能力减弱。通过生物信息学分析和双荧光素酶报告基因测定，我们发现 miR-214 是 HANR 的下游靶基因。此外，miR-214 抑制剂在很大程度上增强了由 HANR 敲低调节的 HCC 细胞的肿瘤表型。HANR 和 miR-214 调节 EZH2，进而影响 TGFBR2 水平。最后，我们证明 EZH2 过表达可以大大挽救 HANR 敲低或 miR-214 模拟诱导的 HCC 肿瘤发生和进展。
在这项研究中，我们报告了一个新发现的调节机制 HANR/miR-214/EZH2/TGF-β 轴，它参与了 HCC 的肿瘤发生和发展。我们的研究结果表明，通过靶向 HANR，可以促进治疗策略或诊断标志物的开发。
