Department of Obstetrics and Gynecology, the 5th People's Hospital of Ji'nan, Ji'nan, Shandong Province, P.R. China.
Eur Rev Med Pharmacol Sci. 2020 Mar;24(5):2294-2302. doi: 10.26355/eurrev_202003_20495.
The aim of this study was to determine the expression profile and the underlying mechanism of the long intergenic non-protein coding RNA AL161431.1 in EC (endometrial carcinoma).
In this study, the expression data for the lncRNA AL161431.1 in EC was downloaded from The Cancer Genome Atlas (TCGA) database and used to examine its expression profile. quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot analysis were used to detect gene and protein expression, respectively. A subcellular fractionation assay was used to determine the location of AL161431.1. Cell Counting Kit-8 (CCK-8) and colony formation assays were used to evaluate cellular proliferation. Cell migration and wound healing assays were used to detect the effects on cell migration. RNA pull-down and Luciferase reporter assays were used to confirm the interaction between AL161431.1 and miR-1252-5p.
High expression levels of AL161431.1 were observed in EC patients, tissues, and cells. Loss-of-function experiments validated the carcinogenic role of AL161431.1. Based on the determined cytoplasmic location of AL161431.1, we investigated the ceRNA network and its relation to AL161431.1, miR-1252-5p, and MAPK (mitogen-activated protein kinase) signaling in EC. The molecular mechanism of the interaction between AL161431.1 and miR-1252-5p, and its effects on the MAPK signaling pathway was validated using rescue experiments in Ishikawa cells.
Our novel results indicate that AL161431.1 targets and binds to miR-1252-5p, resulting in the de-repression of MAPK signaling in EC cells. This highlights the potential for AL161431.1 to be targeted as a potent therapeutic strategy in the treatment of EC.
本研究旨在确定长链非编码 RNA AL161431.1 在子宫内膜癌（EC）中的表达谱及其潜在机制。
本研究从癌症基因组图谱（TCGA）数据库中下载了 EC 中 lncRNA AL161431.1 的表达数据，用于检测其表达谱。采用定量实时聚合酶链反应（qRT-PCR）和 Western blot 分析分别检测基因和蛋白表达。采用亚细胞分馏测定法确定 AL161431.1 的位置。采用细胞计数试剂盒-8（CCK-8）和集落形成实验评估细胞增殖。采用细胞迁移和划痕愈合实验检测细胞迁移的影响。采用 RNA 下拉和荧光素酶报告基因实验验证 AL161431.1 与 miR-1252-5p 的相互作用。
在 EC 患者、组织和细胞中观察到 AL161431.1 的高表达水平。失活功能实验验证了 AL161431.1 的致癌作用。基于确定的 AL161431.1 的细胞质位置，我们研究了 EC 中的 ceRNA 网络及其与 AL161431.1、miR-1252-5p 和 MAPK（丝裂原激活蛋白激酶）信号通路的关系。在 Ishikawa 细胞中进行的挽救实验验证了 AL161431.1 与 miR-1252-5p 相互作用及其对 MAPK 信号通路的影响的分子机制。
我们的新结果表明，AL161431.1 靶向并结合 miR-1252-5p，导致 EC 细胞中 MAPK 信号去抑制。这突出了针对 AL161431.1 的潜在治疗策略在 EC 治疗中的潜力。
