Journal Article

Circ-RNF111 aggravates the malignancy of gastric cancer through miR-876-3p-dependent regulation of KLF12

Zhang, Aimin, Wu, Dongping, Wu, Guoxian, Yang, Yinglin

2021

Overview

Background The aberrant expression of circular RNAs (circRNAs) plays vital roles in the advancement of human cancers, including gastric cancer (GC). In this study, the functions of circRNA ring finger protein 111 (circ-RNF111) in GC were investigated. Methods Quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed for the levels of circ-RNF111, microRNA-876-3p (miR-876-3p) and krueppel-like factor 12 (KLF12) mRNA. RNase R assay was conducted for the feature of circ-RNF111. Cell Counting Kit-8 (CCK-8) assay, colony formation assay, wound-healing assay, and transwell assay were applied for cell viability, colony formation, migration, and invasion, respectively. Flow cytometry analysis was used to analyze cell apoptosis and cell cycle process. The glycolysis level was examined using specific commercial kits. Western blot assay was carried out to measure the protein levels of hexokinase 2 (HK-2) and KLF12. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were employed to verify the combination between miR-876-3p and circ-RNF111 or KLF12. Murine xenograft model was constructed for the role of circ-RNF111 in vivo. Immunohistochemistry (IHC) was used for KLF12 level. Results Circ-RNF111 was higher expressed in GC tissues and cells than normal tissues and cells. Silencing of circ-RNF111 restrained cell viability, colony formation, migration, invasion, cell cycle process and glycolysis and induced apoptosis in GC cells in vitro. Circ-RNF111 positively regulated KLF12 expression via absorbing miR-876-3p. MiR-876-3p downregulation reversed the impacts of circ-RNF111 silencing on GC cell malignant phenotypes. MiR-876-3p overexpression repressed GC cell growth, metastasis and glycolysis, inhibited apoptosis and arrested cell cycle, while KLF12 elevation weakened the effects. Besides, circ-RNF111 knockdown inhibited tumor growth in vivo. Conclusion Circ-RNF111 knockdown relieved the development of GC by regulating miR-876-3p/KLF12 axis.

• Publisher: BioMed Central,BioMed Central Ltd,BMC
• Subject: Analysis / Animals / Antibodies / Apoptosis / Assaying / Binding sites / Cancer / Care and treatment / Cell cycle / Cell Line, Tumor / Cell migration / Cell Movement / Cell Proliferation / Cell viability / Cholecystokinin / Circ-RNF111 / Colonies / Diagnosis / Flow cytometry / Gastric cancer / Gene expression / Glycolysis / Hexokinase / Humans / Immunohistochemistry / Immunoprecipitation / KLF12 / Krueppel-like factor / Kruppel-Like Transcription Factors - genetics / Laboratory animals / Malignancy / Medicine / Medicine & Public Health / Metastases / Mice / MicroRNA / MicroRNAs / MicroRNAs - genetics / miR-876-3p / miRNA / mRNA / Nuclear Proteins / Phenotypes / Polymerase chain reaction / Prognosis / Proteins / Ribonuclease / Ribonucleic acid / RING finger proteins / RNA / RNA, Circular - genetics / RNA, Circular - metabolism / Stomach cancer / Stomach Neoplasms - genetics / Stomach Neoplasms - pathology / Surgical Oncology / Tumors / Ubiquitin-Protein Ligases / Wound healing / Xenografts / Xenotransplantation