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环状RNA-0072309通过靶向微小RNA-665对Hep3B细胞系产生抗肿瘤影响。

Circular RNA-0072309 has antitumor influences in Hep3B cell line by targeting microRNA-665.

作者信息

Yu Qiuyun, Dai Jinhua, Shu Ming

机构信息

Department of Clinical Laboratory, Hwa Mei Hospital, University of Chinese Academy of Science (Ningbo No.2 Hospital), Ningbo, Zhejiang, China.

Department of Hepatobiliary Surgery, Hwa Mei Hospital, University of Chinese Academy of Science (Ningbo No.2 Hospital), Ningbo, Zhejiang, China.

出版信息

Biofactors. 2023 Jan;49(1):79-89. doi: 10.1002/biof.1618. Epub 2020 Feb 11.

Abstract

Liver cancer is a malignant tumor that occurs in the liver and has a high mortality rate. We strived to detect the role and mechanism of circRNA-0072309 in liver cancer. Hep3B cell line was transfected with pc-circ and si-circ for viability, colony formation, apoptosis, migration, and invasion tests, which were individually performed by CCK-8, colony formation detection, flow cytometry assay, migration and invasion assays. What is more, the luciferase reporter assay was conducted to determine the target relationship between the circRNA-0072309 and microRNA (miR)-665. The expression of circRNA-0072309 was examined by qRT-PCR. The expression of proteins was examined via western blot. CircRNA-0072309 was lowly expressed in liver cancer tissues and positively associated with 5-year survival rate. The viability, colony formation, invasive and migratory ability were inhibited by abundant circRNA-0072309, which promoted cell apoptosis on the contrary. CircRNA-0072309 knockdown induced opposite effects, but could not affect apoptosis. Overexpressed miR-665 in tumor tissues was targeted and negatively controlled by circRNA-0072309. The PI3K/AKT and Wnt/β-catenin pathways were inhibited by abundant circRNA-0072309. miR-665 overexpression disturbed those effects derived from pc-circ. The circRNA-0072309 had antitumor influences in Hep3B cell line through targeting miR-665 relying on the deactivation of PI3K/AKT and Wnt/β-catenin pathways.

摘要

肝癌是一种发生于肝脏的恶性肿瘤,死亡率很高。我们致力于探究环状RNA-0072309在肝癌中的作用及机制。用pc-circ和si-circ转染Hep3B细胞系,进行细胞活力、集落形成、凋亡、迁移和侵袭试验,分别通过CCK-8法、集落形成检测、流式细胞术分析、迁移和侵袭试验来完成。此外,进行荧光素酶报告基因检测以确定环状RNA-0072309与微小RNA(miR)-665之间的靶向关系。通过qRT-PCR检测环状RNA-0072309的表达。通过蛋白质印迹法检测蛋白质表达。环状RNA-0072309在肝癌组织中低表达,且与5年生存率呈正相关。大量的环状RNA-0072309可抑制细胞活力、集落形成、侵袭和迁移能力,相反却促进细胞凋亡。敲低环状RNA-0072309则产生相反的效果,但不影响细胞凋亡。肿瘤组织中过表达的miR-665受环状RNA-0072309靶向并受到其负调控。大量的环状RNA-0072309可抑制PI3K/AKT和Wnt/β-连环蛋白信号通路。miR-665过表达会干扰pc-circ产生的这些效应。环状RNA-0072309通过靶向miR-665,依赖PI3K/AKT和Wnt/β-连环蛋白信号通路的失活,对Hep3B细胞系产生抗肿瘤影响。

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