Effect of circRNA hsa_circ_0091579 on the proliferation, migration, and invasion of hepatoma cells

Weikai YU; Tingting FENG; Xiaobing CHEN; Jiye LUO; Wanwen FENG; Yanli WANG

doi:10.3969/j.issn.1001-5256.2021.05.029

Volume 37 Issue 5

May 2021

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Article Navigation > Journal of Clinical Hepatology > 2021 > 37(5): 1126-1131

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Effect of circRNA hsa_circ_0091579 on the proliferation, migration, and invasion of hepatoma cells

DOI: 10.3969/j.issn.1001-5256.2021.05.029

1.
Department of Emergency Medicine, Lianyungang First People's Hospital, Lianyungang, Jiangsu 222000, China
2.
Department of Orthopedics, Lianyungang Municipal Oriental Hospital, Lianyungang, Jiangsu 222042, China

Received Date: 2020-10-15
Accepted Date: 2020-12-10
Published Date: 2021-05-20

Abstract

Abstract

Objective To investigate the expression of circular RNA (cirRNA) hsa_circ_0091579 in human hepatocellular carcinoma (HCC) cell lines and its effect on the proliferation, migration, and invasion of HCC cells. Methods Human HCC cell lines SMMC-7721, Huh-7, MHCC-97H, and HepG2 and normal human liver cell line HL-7702 were cultured in vitro. RNA was extracted from cells and quantitative real-time PCR (qRT-PCR) was used to measure the expression of hsa_circ_0091579 in HCC cell lines and normal human liver cell line. Small interfering RNA (siRNA) was designed for the cyclic splicing site of hsa_circ_0091579, and HepG2 and Huh-7 cells were transfected with hsa_circ_0091579 siRNA in vitro; qRT-PCR was used to verify transfection efficiency. The experiment was divided into siRNA group (transfected with hsa_circ_0091579 siRNA) and NC group (transfected with negative control siRNA), and CCK-8 assay, flow cytometry, wound healing assay, and Transwell assay were used to investigate the effect of hsa_circ_0091579 on cell proliferation, apoptosis, migration, and invasion in HepG2 and Huh-7 cells. Dual luciferase reporter gene assay was used to verify the predicted target. The t-test was used for comparison of continuous data between the two groups. Results Compared with the normal human liver cell line HL-7702, the HCC cell lines SMMC-7721, Huh-7, MHCC-97H, and HepG2 had a significant increase in the expression level of hsa_circ_0091579 (t=14.27, 36.34, 26.70, and 36.16, all P < 0.001). Compared with the NC group, hsa_circ_0091579 siRNA effectively silenced hsa_circ_0091579 in HepG2 and Huh-7 cells (t=14.22 and 27.20, P=0.005 and 0.001). CCK-8 assay and flow cytometry showed that compared with the NC group, the siRNA group had a significant reduction in the proliferative activity of HepG2 and Huh-7 cells and a significant increase in apoptosis rate (all P < 0.05); wound healing assay and Transwell assay showed that compared with the NC group, the siRNA group had significant reductions in the migration and invasion abilities of HepG2 and Huh-7 cells (t=19.63, 13.61, 20.75, and 18.45, P=0.003, 0.005, 0.002, and 0.003). Luciferase reporter assay showed that compared with the NC group, miR-149, miR-490-5p, and miR-502-5p significantly reduced the activity of wild-type luciferase plasmids (t=10.01, 9.13, and 61.49, P=0.010, P=0.012, and P < 0.001). Conclusion Hsa_circ_0091579 is highly expressed in HCC cell lines and may play the role of oncogene by inhibiting miR-149, miR-490-5p, and miR-502-5p.
- Carcinoma, Hepatocellular,
- DNA, Circular,
- Cell Physiological Phenomena

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Effect of circRNA hsa_circ_0091579 on the proliferation, migration, and invasion of hepatoma cells

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Effect of circRNA hsa_circ_0091579 on the proliferation, migration, and invasion of hepatoma cells

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