Proctology Department, The First Affiliated Hospital of Jiamusi University, No.348 Dexiang Street, Jiamusi, 154002, Heilongjiang, China.
General Surgical Department, The First Affiliated Hospital of Jiamusi University, Jiamusi, 154002, Heilongjiang, China.
Dig Dis Sci. 2022 Aug;67(8):3714-3724. doi: 10.1007/s10620-021-07263-0. Epub 2021 Oct 16.
Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide. MicroRNAs (miRNAs) have been widely validated as potential biomarkers for cancer treatment and diagnosis.
This paper intends to study the effect and specific mechanism of miR-574-3p/CUL2 axis in GC.
The miR-574-3p expression in GC tissues and cell lines was analyzed by reverse transcription polymerase chain reaction (RT-PCR). GC cell (N87) proliferation, migration and invasion were determined by the Brdu assay and Transwell assay, respectively. The tumor xenotransplantation model was established in vivo to test the effect of miR-574-3p or Cullin 2 (CUL2) on tumor growth. The relationship between miR-574-3p and CUL2 was predicated by bioinformatic analysis and verified by dual-luciferase reporter assay and RIP experiment. The expression of CUL2, hypoxia-induced transcription factor-1α (HIF-1α) as well as E-cadherin, Snail and Vimentin was monitored by western blot and immunohistochemistry.
miR-574-3p was overexpressed in GC tissues and cells. Forced upregulation of miR-574-3p enhanced proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of GC cells (N87), while downregulation of miR-574-3p resulted in reverse effects. Additionally, miR-574-3p promoted N87 cells growth and EMT in vivo. CUL2 was negatively regulated by miR-574-3p in N87 cells, and upregulation of CUL2 repressed the malignant behaviors of N87 cells. Moreover, CUL2 directly interacted with HIF-1α and suppressed HIF-1α expression both in vitro and in vivo.
miR-574-3p targeted CUL2 to upregulate HIF-1α, thus facilitating the progression of GC.
胃癌（GC）是全球癌症相关死亡的第三大主要原因。microRNAs（miRNAs）已被广泛验证为癌症治疗和诊断的潜在生物标志物。
本研究旨在探讨 miR-574-3p/CUL2 轴在 GC 中的作用及具体机制。
采用逆转录聚合酶链反应（RT-PCR）分析 GC 组织和细胞系中的 miR-574-3p 表达。通过 Brdu 检测和 Transwell 检测分别确定 GC 细胞（N87）的增殖、迁移和侵袭能力。体内建立肿瘤异种移植模型，以检测 miR-574-3p 或 Cullin 2（CUL2）对肿瘤生长的影响。通过生物信息学分析预测 miR-574-3p 与 CUL2 之间的关系，并通过双荧光素酶报告基因检测和 RIP 实验进行验证。通过 Western blot 和免疫组化检测 CUL2、缺氧诱导转录因子-1α（HIF-1α）以及 E-钙黏蛋白、Snail 和波形蛋白的表达。
miR-574-3p 在 GC 组织和细胞中高表达。强制上调 miR-574-3p 增强了 GC 细胞（N87）的增殖、迁移、侵袭和上皮间质转化（EMT），而下调 miR-574-3p 则产生相反的效果。此外，miR-574-3p 促进了 N87 细胞在体内的生长和 EMT。在 N87 细胞中，CUL2 受 miR-574-3p 的负调控，上调 CUL2 抑制了 N87 细胞的恶性行为。此外，CUL2 直接与 HIF-1α 相互作用，在体外和体内均抑制 HIF-1α 的表达。
miR-574-3p 通过靶向 CUL2 上调 HIF-1α，从而促进 GC 的进展。
