Department of neurology, The Affiliated YanAn Hospital of Kunming Medical University, Kunming, 650051, Yunnan, China; Key Laboratory of Tumor Immunological Prevention and Treatment of Yunnan Province, China.
Department of Gastroenterology, The Affiliated YanAn Hospital of Kunming Medical University, Kunming, 650051, Yunnan, China; Key Laboratory of Tumor Immunological Prevention and Treatment of Yunnan Province, China.
Life Sci. 2019 Jan 15;217:271-282. doi: 10.1016/j.lfs.2018.12.024. Epub 2018 Dec 14.
The oncogenic role of lncRNA Hotair has been acknowledged in subset of malignancies, including gastric cancer (GC). However, the detailed molecular mechanisms that contribute to its oncogenic role of are largely elusive. This study was designed to explore the underlying mechanism that contributes the regulatory role of Hotair in GC pathogenesis and progression.
Expression pattern of lncRNAs in GC tissues and adjacent normal tissues were identified by using microarray analysis. The cell proliferation of GC cells was examined by CCK-8 assay and colony formation assay, while migration and invasion capabilities of GC cells were examined by Transwell (with or without Matrigel) assay. Cell apoptosis was examined by Flow cytometer. qRT-PCR and western blotting were used to examine the expression of Hotair, miR-217, and other related genes. The potential target relationships were predicted by miRcode algorithm, and validated by dual luciferase reporter gene assay.
We observed that Hotair was frequently up-regulated in GC tissues and cell lines, and high Hotair level was positively correlated with poor prognosis in GC patients. Knockdown of Hotair inhibited GC cells' viability, migration, invasion, Epithelial mesenchymal transition process. MiR-217 was decreased while GCP5 was increased in GC cells. Hotair negatively regulated the expression of miR-217 in GC while miR-217 targeted GCP5 to down-regulate its expression. Hotair promoted GC development by promoting GCP5 expression via sponging miR-217.
Hotair could serve as a potentially prognostic indicator and provide new light into its underlying biological-molecular mechanism in GC.
长链非编码 RNA Hotair 在包括胃癌（GC）在内的部分恶性肿瘤中具有致癌作用。然而，其致癌作用的详细分子机制在很大程度上仍不清楚。本研究旨在探讨 Hotair 在 GC 发病机制和进展中发挥调节作用的潜在机制。
通过微阵列分析鉴定 GC 组织和相邻正常组织中 lncRNA 的表达模式。通过 CCK-8 测定和集落形成测定检测 GC 细胞的增殖，通过 Transwell（有或没有 Matrigel）测定检测 GC 细胞的迁移和侵袭能力。通过流式细胞仪检测细胞凋亡。qRT-PCR 和 Western blot 用于检测 Hotair、miR-217 和其他相关基因的表达。miRcode 算法预测潜在的靶标关系，并通过双荧光素酶报告基因测定进行验证。
我们观察到 Hotair 在 GC 组织和细胞系中频繁上调，并且 Hotair 水平高与 GC 患者的预后不良呈正相关。Hotair 敲低抑制 GC 细胞的活力、迁移、侵袭和上皮间质转化过程。GC 细胞中 miR-217 减少，GCP5 增加。Hotair 在 GC 中负调控 miR-217 的表达，而 miR-217 通过靶向 GCP5 下调其表达。Hotair 通过海绵吸附 miR-217 促进 GCP5 表达，从而促进 GC 的发展。
Hotair 可以作为一种潜在的预后指标，并为其在 GC 中的潜在生物学-分子机制提供新的见解。
