Nan fang yi ke da xue xue bao = Journal of Southern Medical University · 2021
[Effect of fatty acid synthase gene silencing on lipid metabolism and biological behaviors of human hepatoblastoma HepG2 cells].
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Huang H, Wei Y, Wang J, Ran F, Chen Q
Аннотация
OBJECTIVE: To investigate the effect of fatty acid synthase (FASN) gene silencing on lipid metabolism and biological behaviors of human hepatoblastoma HepG2 cells.
OBJECTIVE: Small interfering RNA (siRNA) targeting FASN gene or a negative control siRNA sequence (NC-siRNA) was transfected into HepG2 cells, and the gene silencing efficiency was evaluated with qRT-PCR and Western blotting. Triglyceride level in the cells was detected using enzyme-linked immunosorbent assay, and Oil red O staining was performed to examine intracellular lipid droplets. The proliferation ability of the transfected cells was tested by CCK-8 assay, and cell apoptosis was evaluated using annexin V-FITC/PI apoptosis detection kit. Wound healing assay and Transwell assay were performed to assess the migration ability of the transfected cells.
OBJECTIVE: Transfection of the cells with FASN-siRNA, but not NC-siRNA, significantly lowered FASN expression at both the mRNA and protein level (P < 0.001) and decreased the number of lipid droplets (P < 0.001) and triglyceride level (P < 0.01) in the cells. FASN gene silencing significantly inhibited proliferation, increased apoptosis rate and suppressed migration of HepG2 cells (P < 0.001).
OBJECTIVE: FASN gene silencing inhibits proliferation and migration and promotes apoptosis of HepG2 cells possibly by suppressing lipid synthesis in the cells.
OBJECTIVE: Small interfering RNA (siRNA) targeting FASN gene or a negative control siRNA sequence (NC-siRNA) was transfected into HepG2 cells, and the gene silencing efficiency was evaluated with qRT-PCR and Western blotting. Triglyceride level in the cells was detected using enzyme-linked immunosorbent assay, and Oil red O staining was performed to examine intracellular lipid droplets. The proliferation ability of the transfected cells was tested by CCK-8 assay, and cell apoptosis was evaluated using annexin V-FITC/PI apoptosis detection kit. Wound healing assay and Transwell assay were performed to assess the migration ability of the transfected cells.
OBJECTIVE: Transfection of the cells with FASN-siRNA, but not NC-siRNA, significantly lowered FASN expression at both the mRNA and protein level (P < 0.001) and decreased the number of lipid droplets (P < 0.001) and triglyceride level (P < 0.01) in the cells. FASN gene silencing significantly inhibited proliferation, increased apoptosis rate and suppressed migration of HepG2 cells (P < 0.001).
OBJECTIVE: FASN gene silencing inhibits proliferation and migration and promotes apoptosis of HepG2 cells possibly by suppressing lipid synthesis in the cells.