Department of Urology, Dongying People's Hospital, Dongying, Shandong, China.
Eur Rev Med Pharmacol Sci. 2018 Oct;22(20):6640-6649. doi: 10.26355/eurrev_201810_16139.
To explore the role of HOTAIR in the pathogenesis of adrenocortical carcinoma (ACC) and its underlying mechanism.
Differentially expressed lncRNA (HOTAIR) in ACC was screened out from the GEO database. The survival analysis and ROC curve were performed according to HOTAIR expressions in ACC patients. The correlation between HOTAIR expression and clinical information of ACC patients was analyzed by chi-square test. The univariate and multivariate COX regression analysis was carried out to analyze the relationship between HOTAIR expression, disease-free survival (DFS) and overall survival (OS) of ACC patients. We then detected HOTAIR expression in 77 ACC tissues and 30 normal tissues by qRT-PCR (quantitative Real-time polymerase chain reaction). ACC cell lines were further screened out for the following in vitro experiments. After altering HOTAIR expression in ACC cells by plasmid transfection, proliferation and cell cycle were detected by Cell Counting Kit-8 (CCK-8) and colony formation assay, respectively. Finally, Western blot was utilized to detect expressions of cell cycle-related genes in ACC cells.
HOTAIR was overexpressed in ACC tissues than that of normal tissues. HOTAIR expression was remarkably increased in ACC with T3 and T4 stage than that of T1 and T2 stage. Moreover, HOTAIR expression was remarkably increased in ACC with stage III and IV than that of stage I and II. HOTAIR was an independent prognostic factor for DFS and OS of ACC patients. For in vitro experiments, inhibited proliferation and arrested cell cycle were observed in H295R cells transfected with si-HOTAIR. Opposite results were obtained after SW-13 cells were transfected with HOTAIR overexpression plasmid. Furthermore, expressions of cell cycle-related genes, including Cyclin D1, p-Rb and p-GSK3β were remarkably decreased after HOTAIR knockdown.
We demonstrated for the first time that HOTAIR is overexpressed in ACC and is a prognostic risk factor in ACC patients. HOTAIR participates in the development and progression of ACC via shortening cell cycle and promoting proliferation of ACC cells.
探讨长链非编码 RNA（HOTAIR）在肾上腺皮质癌（ACC）发病机制中的作用及其潜在机制。
从 GEO 数据库中筛选出 ACC 中差异表达的 lncRNA（HOTAIR）。根据 ACC 患者 HOTAIR 表达情况进行生存分析和 ROC 曲线分析。采用卡方检验分析 HOTAIR 表达与 ACC 患者临床资料的相关性。采用单因素和多因素 COX 回归分析 HOTAIR 表达与 ACC 患者无病生存率（DFS）和总生存率（OS）的关系。然后采用 qRT-PCR（实时定量聚合酶链反应）检测 77 例 ACC 组织和 30 例正常组织中的 HOTAIR 表达。进一步筛选出 ACC 细胞系进行以下体外实验。通过质粒转染改变 ACC 细胞中的 HOTAIR 表达后，通过细胞计数试剂盒-8（CCK-8）和集落形成实验分别检测细胞增殖和细胞周期。最后，通过 Western blot 检测 ACC 细胞中细胞周期相关基因的表达。
HOTAIR 在 ACC 组织中的表达高于正常组织。与 T1 期和 T2 期相比，T3 期和 T4 期 ACC 中 HOTAIR 的表达显著增加。此外，与 I 期和 II 期相比，III 期和 IV 期 ACC 中 HOTAIR 的表达显著增加。HOTAIR 是 ACC 患者 DFS 和 OS 的独立预后因素。在体外实验中，转染 si-HOTAIR 的 H295R 细胞增殖受到抑制，细胞周期停滞。转染 HOTAIR 过表达质粒的 SW-13 细胞则获得相反的结果。此外，下调 HOTAIR 表达后，细胞周期相关基因的表达，包括 Cyclin D1、p-Rb 和 p-GSK3β，明显降低。
我们首次证明 HOTAIR 在 ACC 中过表达，是 ACC 患者的预后危险因素。HOTAIR 通过缩短细胞周期和促进 ACC 细胞增殖参与 ACC 的发生和发展。
