Department of translational Medical Center, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou, China.
Department of Medicine, University of California, San Francisco, San Francisco, CA, USA.
Cancer Med. 2023 Jan;12(1):445-458. doi: 10.1002/cam4.4915. Epub 2022 Jun 2.
Long non-coding RNAs (lncRNAs) have critical functions within esophageal squamous cell carcinoma (ESCC). However, the function and mechanism underlying ESCC-associated lncRNA-1 (ESCCAL-1) in ESCC tumorigenesis have not been well clarified.
ESCCAL-1, miR-590 and LRP6 were quantified using qRT-PCR. Cell viability, migration and invasion abilities were measured using CCK-8 assay and transwell assays. The protein pression was determined with western blot assay. The xenograft model assays were used to examine the impact of ESCCAL-1 on tumorigenic effect in vivo. Direct relationships among ESCCAL-1, miR-590 and LRP6 were confirmed using dual-luciferase reporter assays.
The present work discovered the ESCCAL-1 up-regulation within ESCC. Furthermore, ESCCAL-1 was found to interact with miR-590 and consequently restrict its expression. Functionally, knocking down ESCCAL-1 or over-expressing miR-590 hindered ESCC cell growth, invasion, and migration in vitro. Moreover, inhibition of miR-590 could reverse the effect of knockdown of ESCCAL-1 on cells. Importantly, it was confirmed that LRP6 was miR-590's downstream target and LRP6 over-expression also partly abolished the role of miR-590 overexpression in ESCC cells.
We have uncovered a novel regulatory network comprising aberrant interaction of ESCCAL-1/miR-590/LRP6 participated in ESCC progression.
长链非编码 RNA（lncRNA）在食管鳞状细胞癌（ESCC）中具有重要功能。然而，ESCC 相关 lncRNA-1（ESCCAL-1）在 ESCC 肿瘤发生中的功能和机制尚未得到充分阐明。
使用 qRT-PCR 定量检测 ESCCAL-1、miR-590 和 LRP6。使用 CCK-8 测定法和 Transwell 测定法测定细胞活力、迁移和侵袭能力。使用 Western blot 测定法测定蛋白质表达。使用异种移植模型测定法研究 ESCCAL-1 对体内致瘤作用的影响。使用双荧光素酶报告基因测定法证实 ESCCAL-1、miR-590 和 LRP6 之间的直接关系。
本研究发现 ESCC 中 ESCCAL-1 的上调。此外，发现 ESCCAL-1 与 miR-590 相互作用，从而限制其表达。功能上，敲低 ESCCAL-1 或过表达 miR-590 可抑制 ESCC 细胞在体外的生长、侵袭和迁移。此外，抑制 miR-590 可逆转敲低 ESCCAL-1 对细胞的作用。重要的是，证实 LRP6 是 miR-590 的下游靶标，LRP6 过表达也部分消除了 miR-590 过表达在 ESCC 细胞中的作用。
我们发现了一个由 ESCCAL-1/miR-590/LRP6 异常相互作用组成的新型调控网络，参与 ESCC 的进展。
