Yang Luoluo, Wang Min, He Ping
Department of Gastroenterology, First Hospital of Jilin University, Changchun 130021, Jilin, People's Republic of China.
Department of Pathology, Jilin Provincial Cancer Hospital, Changchun 130012, Jilin, People's Republic of China.
Cancer Manag Res. 2020 Nov 19;12:11845-11855. doi: 10.2147/CMAR.S267666. eCollection 2020.
The therapy of patients with advanced phase gastric cancer is still a huge threat, with extremely imperfect therapies authorized. Even if the amassed indications have validated the significance of lncRNA in gastric cancer, few understandings are stated concerning nuclear paraspeckle assembly transcript 1 (NEAT1) practical functions and molecular mechanisms.
In this research, the expression of NEAT1 and miR-1224-5p in gastric cancer tissues was measured by qRT-PCR analysis, and the expression of remodeling and spacing factor 1 (RSF1) was measured by IHC assay. Then, the bioinformatics prediction software ENCORI was applied to envisage the assumed binding sites. The monitoring roles of NEAT1 or miR-1224-5p on the cell proliferation and migration capacity were verified by CCK-8, wound healing and transwell assay, correspondingly. The interactions among NEAT1, miR-1224-5p and RSF1 were investigated via luciferase analysis.
Our findings revealed high expression levels of NEAT1, RSF1 and a decreased expression level of miR-1224-5p in gastric cancer. Upregulation of NEAT1 or knockdown of miR-1224-5p elevated gastric cancer cell proliferation, and migration. Bioinformatics and luciferase analyses simplified that NEAT1 directly cooperated with miR-1224-5p to weaken miR-1224-5p binding to the RSF1 3'-UTR region. Likewise, the mechanical inquiries ratified that initiation of the miR-1224-5p/RSF1 regulatory loop by miR-1224-5p knockdown or overexpressed RSF1 validated the functions of NEAT1 in endorsing gastric cancer cell malignancy.
Our research initially validated that NEAT1 may regulate the expression of RSF1 competitive sponge to miR-1224-5p, contributed to the supervision of gastric cancer evolution, which exposed new brightness for diagnosis and therapy of gastric cancer.
晚期胃癌患者的治疗仍然是一个巨大的挑战，目前获批的治疗方法极不完善。尽管大量研究表明长链非编码RNA（lncRNA）在胃癌中具有重要意义，但关于核旁斑组装转录本1（NEAT1）的实际功能和分子机制，人们了解甚少。
在本研究中，通过qRT-PCR分析检测胃癌组织中NEAT1和miR-1224-5p的表达，通过免疫组化检测重塑和间距因子1（RSF1）的表达。然后，应用生物信息学预测软件ENCORI预测假定的结合位点。分别通过CCK-8、伤口愈合实验和Transwell实验验证NEAT1或miR-1224-5p对细胞增殖和迁移能力的影响。通过荧光素酶分析研究NEAT1、miR-1224-5p和RSF1之间的相互作用。
我们的研究结果显示，胃癌组织中NEAT1和RSF1表达水平较高，而miR-1224-5p表达水平降低。上调NEAT1或敲低miR-1224-5p可提高胃癌细胞的增殖和迁移能力。生物信息学和荧光素酶分析表明，NEAT1直接与miR-1224-5p相互作用，削弱miR-1224-5p与RSF1 3'-UTR区域的结合。同样，机制研究证实，敲低miR-1224-5p或过表达RSF1启动miR-1224-5p/RSF1调控环，验证了NEAT1在促进胃癌细胞恶性增殖中的作用。
我们的研究首次证实，NEAT1可能通过竞争性结合miR-1224-5p来调节RSF1的表达，促进胃癌进展，为胃癌的诊断和治疗提供了新的思路。
