H Huang1, Y Wei1, J Wang1, F Ran1, Q Chen1. 1. Department of Experiment Center, Dongfeng Hospital Affiliated to Hubei University of Medicine, Shiyan 442008, China.
Abstract
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: 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.
Authors: Nousheen Zaidi; Leslie Lupien; Nancy B Kuemmerle; William B Kinlaw; Johannes V Swinnen; Karine Smans Journal: Prog Lipid Res Date: 2013-08-31 Impact factor: 16.195
Authors: Yekaterina Y Zaytseva; Victoria A Elliott; Piotr Rychahou; W Conan Mustain; Ji Tae Kim; Joseph Valentino; Tianyan Gao; Kathleen L O'Connor; Janna M Neltner; Eun Y Lee; Heidi L Weiss; B Mark Evers Journal: Carcinogenesis Date: 2014-02-07 Impact factor: 4.944