Zheng Mei, Liu Xingguang, Zhou Qin, Liu Gangli
Department of Traditional Chinese Medicine, Qianfoshan Hospital Affiliated to Shandong University, Ji'nan, 250014, Shandong, China.
Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Shandong University, Ji'nan, 250012, Shandong, China.
Cancer Med. 2018 Jul;7(7):3143-3156. doi: 10.1002/cam4.1523. Epub 2018 Jun 14.
This study is aimed to explore the regulatory effect of lncRNA HOTAIR/miR-148a/DLGAP1 axis on head and neck tumor (HNT) cell growth, cell mobility, and invasiveness. HOTAIRM1, miR-148a, and DLGAP1 level in HNT tissues and adjacent normal tissues were measured by qRT-PCR. Cell Counting Kit-8 (CCK-8) and Transwell (migration and invasion) assay were used to survey the influence of HOTAIRM1, miR-148a, and DLGAP1 on Fadu cells. Nude mouse xenograft was utilized to validate the influence of HOTAIRM1 in vivo. Dual-luciferase reporter assay confirms the relationship between HOTAIRM1 and miR-148a, miR-148a, and DLGAP1. The expression level of HOTAIRM1 was downregulated in human HNT tissues and cells. Overexpression of HOTAIRM1 significantly moderated Fadu cells proliferation, apoptosis, migration, and invasion in vitro and impaired the tumorigenesis in vivo. The expression level of miR-148a was upregulated in human HNT tissue compared to the adjacent tissues. We identified that miR-148a was a target of HOTAIRM1 and its expression levels were reduced by HOTAIRM1. Transfection of miR-148a mimics increased proliferation, migration, and invasion of Fadu cells. DLGAP1 was identified as a novel target of miR-148a and its expression level was promoted by either HOTAIRM1 overexpression or miR-148a knockdown. Overexpression of DLGAP1 also facilitated the cell viability and metastasis of Fadu cells. HOTAIRM1 was confirmed as a tumor suppressor via sponging miR-148a and promote the expression of DLGAP1, which could be regarded as an important target for the prevention and treatment of HNT.
本研究旨在探讨长链非编码RNA HOTAIR/miR-148a/DLGAP1轴对头颈部肿瘤（HNT）细胞生长、细胞迁移和侵袭的调控作用。采用qRT-PCR检测HNT组织及癌旁正常组织中HOTAIRM1、miR-148a和DLGAP1的水平。使用细胞计数试剂盒-8（CCK-8）和Transwell（迁移和侵袭）实验来研究HOTAIRM1、miR-148a和DLGAP1对Fadu细胞的影响。利用裸鼠异种移植模型在体内验证HOTAIRM1的作用。双荧光素酶报告基因实验证实了HOTAIRM1与miR-148a、miR-148a与DLGAP1之间的关系。HOTAIRM1在人HNT组织和细胞中的表达水平下调。HOTAIRM1的过表达显著抑制了Fadu细胞在体外的增殖、凋亡、迁移和侵袭，并在体内抑制了肿瘤发生。与癌旁组织相比，人HNT组织中miR-148a的表达水平上调。我们发现miR-148a是HOTAIRM1的靶标，且其表达水平受到HOTAIRM1的下调。转染miR-148a模拟物可增加Fadu细胞的增殖、迁移和侵袭。DLGAP1被鉴定为miR-148a的新靶标，其表达水平可通过HOTAIRM1过表达或miR-148a敲低而升高。DLGAP1的过表达也促进了Fadu细胞的活力和转移。HOTAIRM1通过吸附miR-148a并促进DLGAP1的表达而被证实为一种肿瘤抑制因子，这可被视为HNT防治的重要靶点。
