Targeting a novel circITCH/miR-421/BTG1 axis is effective to suppress the malignant phenotypes in hepatocellular carcinoma (HCC) cells.

Abstract

Circular RNA-based competing endogenous RNA (ceRNA) networks contribute to the initiation and development of various types of cancer, including hepatocellular carcinoma (HCC). Although a novel circular RNA itchy E3 ubiquitin protein ligase (circITCH) is identified as a tumor suppressor in HCC, its detailed molecular mechanisms have not been fully delineated. The present study was designed to resolve this issue, and we firstly verified that circITCH suppressed the malignant phenotypes in HCC cells by regulating a novel miR-421/B-cell translocation gene 1 (BTG1) axis. Specifically, through performing the Real-Time qPCR analysis, we noticed that circITCH expression in HCC tumor tissues or cell lines were significantly lower than that in adjacent normal tissues or normal hepatocytes, and the expression levels of circITCH were negatively correlated with tumor size and TNM stage in HCC patients. Next, our functional experiments confirmed that overexpression of circITCH induced cell cycle arrest and apoptosis, and reduced cell viability and colony forming ability in Hep3B and Huh7 cells. Mechanically, bioinformatics analysis, RNA immunoprecipitation and luciferase reporter assay demonstrated that circITCH served as RNA sponges for miR-421 to elevate BTG1 levels in HCC cells. The rescuing experiments verified that upregulation of miR-421 promoted cell viability and colony formation, and reduced apoptosis, which were abrogated by overexpression of circITCH or BTG1. In conclusion, this study identified a novel circITCH/miR-421/BTG1 axis that restrained the development of HCC, and our findings provided novel biomarkers for the treatment of this disease.