长链非编码 RNA VIM-AS1 的敲低通过 miR-105-5p 靶向调节 WEE1 抑制神经胶质瘤细胞增殖和迁移，并增加细胞凋亡。
Knockdown of long non-coding RNA VIM-AS1 inhibits glioma cell proliferation and migration, and increases the cell apoptosis via modulation of WEE1 targeted by miR-105-5p.
机构信息
Neurosurgery Department, Shangluo Central Hospital, Shangluo City, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Jun;24(12):6834-6847. doi: 10.26355/eurrev_202006_21673.
OBJECTIVE
Glioma including glioblastoma is the main type of primary brain tumors worldwide. LncRNAs have participated in glioma formation. This study aims to investigate the underlying mechanism for VIM-AS1/miR-105-5p/WEE1 signaling in glioma.
PATIENTS AND METHODS
The clinical tumors and adjacent tissues were collected from 24 patients with glioma in the Shang Luo Central Hospital. Then, the clinical samples were subjected to hematoxylin-eosin staining (H&E). VIM-AS1, miR-105-5p, and WEE1 levels were measured using real-time PCR. The protein levels of WEE1, Cyclin A1, PCNA, N-cadherin, Vimentin, and Bcl-2, E-cadherin, and Bax were analyzed using Western blot. The overall survival of glioma patients was evaluated using the Kaplan-Meier analysis. The interaction between VIM-AS1 and miR-105-5p was determined using RIP assay and Dual-Luciferase reporter assay, and the binding between miR-105-5p and WEE1 was also detected by Dual-Luciferase reporter assay. Cell proliferation, colony formation, cell cycle, apoptosis, and migration were confirmed using CCK-8, colony formation assay, flow cytometry, and transwell assay, respectively.
RESULTS
VIM-AS1 was elevated in cancer tissues, and high level of VIM-AS1 was positively correlated with poor overall survival. Then, VIM-AS1 could bind to and downregulate miR-105-5p. Furthermore, the knockdown of VIM-AS1 significantly suppressed tumor growth in vivo. The knockdown of VIM-AS1/overexpression of miR-105-5p inhibited glioma cell growth, colony formation, and migration, and enhanced the cell apoptosis by inhibiting expression of Cyclin A1, PCNA, Vimentin, N-cadherin, and Bcl-2, and by increasing the expression of Bax and E-cadherin. Interestingly, the overexpression of VIM-AS1 reversed the tumor-suppressing role of miR-105-5p in glioma cells. Besides, the expression of WEE1 was synergistically regulated by VIM-AS1 and miR-105-5p. Consequently, VIM-AS1 promoted glioma progression via upregulating WEE1 or downregulating miR-105-5p.
CONCLUSIONS
VIM-AS1/miR-105-5p/WEE1 signaling may be a promising target for glioma treatment.
目的
胶质瘤包括胶质母细胞瘤是全球原发性脑肿瘤的主要类型。长链非编码 RNA 已参与胶质瘤的形成。本研究旨在探讨 VIM-AS1/miR-105-5p/WEE1 信号在胶质瘤中的潜在机制。
患者和方法
本研究共纳入 24 名在上海罗中央医院接受治疗的胶质瘤患者的临床肿瘤和相邻组织。然后，对临床样本进行苏木精-伊红（H&E）染色。采用实时 PCR 检测 VIM-AS1、miR-105-5p 和 WEE1 水平。采用 Western blot 分析 WEE1、Cyclin A1、PCNA、N-钙粘蛋白、波形蛋白和 Bcl-2、E-钙粘蛋白和 Bax 的蛋白水平。采用 Kaplan-Meier 分析评估胶质瘤患者的总生存率。采用 RIP 测定和双荧光素酶报告基因测定确定 VIM-AS1 与 miR-105-5p 之间的相互作用，采用双荧光素酶报告基因测定检测 miR-105-5p 与 WEE1 之间的结合。采用 CCK-8、集落形成实验、流式细胞术和 Transwell 实验分别确定细胞增殖、集落形成、细胞周期、细胞凋亡和迁移。
结果
VIM-AS1 在癌组织中上调，高水平的 VIM-AS1 与总生存期不良呈正相关。然后，VIM-AS1 可以与 miR-105-5p 结合并下调其表达。此外，VIM-AS1 的敲低显著抑制体内肿瘤生长。VIM-AS1 的敲低/miR-105-5p 的过表达抑制胶质瘤细胞的生长、集落形成和迁移，并通过抑制 Cyclin A1、PCNA、波形蛋白、N-钙粘蛋白和 Bcl-2 的表达，增加 Bax 和 E-钙粘蛋白的表达，增强细胞凋亡。有趣的是，VIM-AS1 的过表达逆转了 miR-105-5p 对胶质瘤细胞的抑瘤作用。此外，WEE1 的表达受到 VIM-AS1 和 miR-105-5p 的协同调节。因此，VIM-AS1 通过上调 WEE1 或下调 miR-105-5p 促进胶质瘤的进展。
结论
VIM-AS1/miR-105-5p/WEE1 信号可能是治疗胶质瘤的有前途的靶点。
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