长链非编码 RNA TOB1-AS1 通过 Wnt/β-catenin 通路调控 miR-23a/NEU1 轴影响胃癌细胞的增殖、凋亡、迁移和侵袭。
Long non-coding RNA TOB1-AS1 modulates cell proliferation, apoptosis, migration and invasion through miR-23a/NEU1 axis via Wnt/b-catenin pathway in gastric cancer.
机构信息
Department of Radiation Oncology, The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an No. 2 Hospital, Jiangsu, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Nov;23(22):9890-9899. doi: 10.26355/eurrev_201911_19554.
OBJECTIVE
Gastric cancer (GC) is the fourth common cancer worldwide. Long non-coding RNA TOB1 antisense RNA 1 (TOB1-AS1) has been found to participate in the process of GC, while the precise role of TOB1-AS1 is still not understood in GC progression.
MATERIALS AND METHODS
We collected 21-paired GC and para-carcinoma tissue specimens, and the levels of TOB1-AS1 and lysosomal sialidase (NEU1) were detected by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). The protein expression levels of NEU1, b-catenin, c-Myc, Cyclin D1, N-cadherin were determined via Western blot. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry was performed to evaluate cell proliferation. Besides, GC cell migration and invasion capacities were identified by transwell assay. Dual-Luciferase reporter assay was employed to examine the interrelation between miR-23a and TOB1-AS1 or NEU1. Finally, the role of TOB1-AS1 was verified in vivo.
RESULTS
The levels of TOB1-AS1 were decreased in GC tissues and cell lines. Either TOB1-AS1 or NEU1 upregulation accelerated GC cell apoptosis, hampered proliferation, migration, and invasion. Further, the role of TOB1-AS1 silencing on cell behaviors was abrogated by NEU1 upregulation. TOB1-AS1 and NEU1 exerted their roles via Wnt/b-catenin signaling pathway. Overexpression of TOB1-AS1 blocked GC development in vivo. Mechanically, miR-23a was targeted by TOB1-AS1, but directly targeted NEU1.
CONCLUSIONS
TOB1-AS1/miR-23a/NEU1 axis regulated proliferation, apoptosis, migration, and invasion of GC cells via Wnt/b-catenin pathway, providing the evidence for serving TOB1-AS1 as an underlying therapeutic target in human GC treatment.
目的
胃癌（GC）是全球第四大常见癌症。长链非编码 RNA TOB1 反义 RNA 1（TOB1-AS1）已被发现参与 GC 过程，但其在 GC 进展中的精确作用仍不清楚。
材料和方法
我们收集了 21 对 GC 和癌旁组织标本，通过实时定量聚合酶链反应（qRT-PCR）检测 TOB1-AS1 和溶酶体唾液酸酶（NEU1）的水平。通过 Western blot 测定 NEU1、β-连环蛋白、c-Myc、细胞周期蛋白 D1、N-钙粘蛋白的蛋白表达水平。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）测定法评估细胞增殖。通过流式细胞术评估细胞增殖。此外，通过 Transwell 测定法鉴定 GC 细胞迁移和侵袭能力。双荧光素酶报告基因检测法用于检测 miR-23a 与 TOB1-AS1 或 NEU1 之间的相互关系。最后，在体内验证了 TOB1-AS1 的作用。
结果
TOB1-AS1 在 GC 组织和细胞系中的水平降低。TOB1-AS1 或 NEU1 的上调加速了 GC 细胞凋亡，抑制了增殖、迁移和侵袭。此外，NEU1 的上调削弱了 TOB1-AS1 沉默对细胞行为的作用。TOB1-AS1 和 NEU1 通过 Wnt/β-连环蛋白信号通路发挥作用。TOB1-AS1 的过表达在体内阻断了 GC 的发展。机制上，TOB1-AS1 靶向 miR-23a，但直接靶向 NEU1。
结论
TOB1-AS1/miR-23a/NEU1 轴通过 Wnt/β-连环蛋白通路调节 GC 细胞的增殖、凋亡、迁移和侵袭，为将 TOB1-AS1 作为人类 GC 治疗的潜在治疗靶点提供了证据。
Zotero 插件