Department of Gastric Surgery, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huai'an City, Jiangsu Province, P.R. China.
Eur Rev Med Pharmacol Sci. 2019 Aug;23(16):6914-6926. doi: 10.26355/eurrev_201908_18731.
To investigate the molecular mechanisms of long noncoding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) in gastric cancer (GC) development progress.
Relative mRNA and protein expression levels were quantified by quantitative Reverse Transcription-PCR (qRT-PCR) or Western blot analysis. Cell proliferation and cell apoptosis were measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and flow cytometry, respectively. Binding sites of miR-497-5p on NEAT1 or phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1) were determined by RNA pull-down assay or dual-luciferase reporter assay. Finally, the tumorigenic role of NEAT1 in GC was assessed using a xenograft model on nude mice.
NEAT1 was upregulated in GC tissues, promoted proliferation, and inhibited apoptosis of GC cells. NEAT1 could directly bind to and negatively regulate miR-497-5p expression. PIK3R1 was then identified as a downstream target of miR-497-5P. In GC cell models, PIK3R1 was found to be directly negatively regulated by miR-497-5p and indirectly positively regulated by NEAT1. Finally, NEAT1 knockdown inhibited tumor growth, increased miR-497-5p expression, and decreased PIK3R1 expression in xenograft model mice compared with the negative control.
Functioned as an oncogene, NEAT1 promoted cell growth in GC by regulating miR-497-5p/PIK3R1 axis. These results provided valuable insights into the underlying regulation signaling in gastric cancer development, shedding light on NEAT1 a promising therapeutic target from bench to clinic.
研究长链非编码 RNA（lncRNA）核斑浆转录物 1（NEAT1）在胃癌（GC）发展进程中的分子机制。
通过定量逆转录-PCR（qRT-PCR）或 Western blot 分析定量相对 mRNA 和蛋白表达水平。通过 MTT（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴盐）测定和流式细胞术分别测量细胞增殖和细胞凋亡。通过 RNA 下拉测定或双荧光素酶报告基因测定确定 miR-497-5p 与 NEAT1 或磷酯酰肌醇-3-激酶调节亚基 1（PIK3R1）的结合位点。最后，通过裸鼠异种移植模型评估 NEAT1 在 GC 中的致瘤作用。
NEAT1 在 GC 组织中上调，促进 GC 细胞增殖并抑制细胞凋亡。NEAT1 可直接结合并负调控 miR-497-5p 的表达。PIK3R1 随后被鉴定为 miR-497-5P 的下游靶标。在 GC 细胞模型中，发现 PIK3R1 直接受 miR-497-5p 负调控，间接受 NEAT1 正调控。最后，与阴性对照相比，NEAT1 敲低抑制了异种移植模型小鼠的肿瘤生长，增加了 miR-497-5p 的表达，并降低了 PIK3R1 的表达。
作为一种癌基因，NEAT1 通过调节 miR-497-5p/PIK3R1 轴促进 GC 细胞生长。这些结果为胃癌发展中潜在的调控信号提供了有价值的见解，表明 NEAT1 是一种有前途的从实验室到临床的治疗靶点。
