Circular RNA ZNF292 affects proliferation and apoptosis of hepatocellular carcinoma cells by regulating Wnt/β-catenin pathway

C.-H. Chen, Y.-J. Su, H. Ding, J. Duan, J. Wang

Department of Hepatobiliary Surgery, The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China. wxjyrmyy@163.com

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• Oncology

OBJECTIVE: The purpose of this study was to explore the function of circular ribonucleic acid (circRNA) zinc finger protein 292 (ZNF292) in hepatocellular carcinoma (HCC).

MATERIALS AND METHODS: The expression of circRNA ZNF292 in Huh-7 cells was knocked down by small interfering RNAs (siRNAs), and the effect of circRNA ZNF292 knockdown on the proliferation of Huh-7 cells was analyzed by Cell Counting Kit-8 (CCK-8) assay and colony formation assay. Then, flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) were adopted to analyze the impacts of circRNA ZNF292 knockdown on the cycle distribution and apoptosis of Huh-7 cells. Besides, the influences of circRNA ZNF292 knockdown on Wnt/β-catenin signaling pathway and its downstream molecules were detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting.

RESULTS: Compared with those in siRNA-normal control (NC) group, the proliferation of Huh-7 cells was significantly inhibited and their cloning ability was remarkably weakened (p<0.05), the proportion of cells in S phase was decreased while that in G1 phase was increased (p<0.05), the apoptosis rate of Huh-7 cells was higher and the number of apoptosis was larger in siRNA-2# knockdown group (p<0.05). Besides, in Huh-7 cells with circRNA ZNF292 knockdown, the expressions of Axin, β-catenin, phosphorylated signal transducer and activator of transcription 3 (p-STAT3), p-STAT5, Cyclin A and Cyclin-dependent kinase 2 (CDK2) were down-regulated, while the expressions of STAT3 and STAT5 did not change remarkably.

CONCLUSIONS: Knock downing circRNA ZNF292 leads to cell cycle arrest in G1 phase, thus suppressing cell proliferation and promoting cell apoptosis. The regulatory mechanism of circRNA ZNF292 may involve the regulation of cell cycle and related genes.

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