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ISSN 1001-5256 (Print)
ISSN 2097-3497 (Online)
CN 22-1108/R
Volume 37 Issue 8
Aug.  2021
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Article Navigation >  Journal of Clinical Hepatology  > 2021  >  37(8): 1873-1877

Effect of small interfering RNA silencing of the AFP gene on the migration and invasion of hepatocellular carcinoma HepG2 cells

DOI: 10.3969/j.issn.1001-5256.2021.08.024

  • Department of General Surgery, Lanzhou University Second Hospital, Lanzhou 730030, China

Research funding:

Gansu Province Science and Technology Foundation (21JR1RA149);

Gansu Province Science and Technology Foundation (21JR1RA161);

Lanzhou Science and Technology Plan Guidance Project (2019-ZD-50);

Lanzhou Science and Technology Plan Guidance Project (2019-ZD-71)

  • Received Date: 2020-12-30
  • Accepted Date: 2021-02-03
  • Published Date: 2021-08-20
    Abstract
  •   Objective  To investigate the effect of alpha-fetoprotein (AFP) on the migration and invasion abilities of hepatocellular carcinoma (HCC) HepG2 cells.  Methods  HCC HepG2 cells were transfected with synthesized small interference RNA (siRNA) for targeted silencing of AFP, and then HepG2 cells were divided into blank control group, negative control group, and AFP siRNA group. After 48 hours of intervention, quantitative real-time PCR and ELISA were used to measure the efficiency of silencing after transfection; Transwell assay was used to analyze the invasion and migration abilities of HepG2 cells after silencing of the AFP gene; Western blot was used to investigate the effect of AFP silencing on the expression of epithelial-mesenchymal transition (EMT)-related proteins (N-cadherin, vimentin, and E-cadherin), AKT, and p-AKT. A one-way analysis of variance was used for comparison of continuous data between groups, and the LSD-t-test was used for further comparison between two groups.  Results  After transfection and silencing, compared with the blank control group, the AFP siRNA group had a significant reduction in the relative mRNA expression of AFP (P < 0.01), with an inhibition rate of 86.440%, and the AFP siRNA group also had a significant reduction in the protein expression level of AFP in cell supernatant (P < 0.01). Compared with the blank control group, the AFP siRNA group had significant reductions in the numbers of migrating and invading cells (both P < 0.01). After the expression of the AFP gene was silenced in HepG2 cells, compared with the blank control group, the AFP siRNA group had a significant increase in the expression level of the EMT-related protein E-cadherin (P < 0.01) and significant reductions in the expression levels of the EMT-related proteins N-cadherin and vimentin (both P < 0.05), as well as a significant reduction in the protein expression level of p-AKT involved in the PI3K/AKT signaling pathway (P < 0.01).  Conclusion  Silencing AFP can inhibit the migration of HepG2 cells, possibly by blocking the PI3K/AKT signaling pathway and inhibiting EMT based on the mechanism of action of HepG2 cell line.

     

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