Experimental study on the effect of Long-chain acyl-CoA synthase 4 on the biological behavior of ovarian cancer cells
MA Rui
WEI Bin
GUAN Xiaodong
JI Xiaoxia
QIN Qian
YANG Cong
SHANG Yun
Abstract：Objective To investigate the expression of acyl-CoA synthase long chain family member 4(ACSL4)in ovarian cancer tissues and its relationship with the biological behavior of ovarian cancer cells.Methods The expression of ACSL4 in ovarian cancer tissues was analyzed by TCGA and GTEx databases,and survival analysis was performed using the Kaplan-Meier Plotter database.The ovarian cancer SKOV-3 cell line was cultured in vitro.Ferroptosis was induced by the ferroptosis activator(Erastin),and the lipid peroxidation level,cell viability,migration and invasion abilities of SKOV-3 cells were detected.The ACSL4 overexpression cell line(oe-ACSL4)was constructed to further verify its functions in ferroptosis and cell biological behaviors.Results Compared with normal tissues,the expression of ACSL4 in TCGA-OV tissues was significantly decreased(P=0.029).The median overall survival time(OS)and median progression-free survival time(PFS)of OV patients with high expression of ACSL4 were significantly better than those of the low expression group(OS:62.34 months vs.59.85 months,P=0.048;)PFS:31.68 months vs.26.14 months,P=0.008).In serous ovarian cancer,the OS in the ACSL4 high-expression group was also significantly prolonged(51.36 months vs.45.78 months,P=0.016).In the Erastin group,SKOV-3 cells significantly upregulated the expression of ACSL4(protein:0.42±0.18 vs.0.15±0.09,P=0.017;mRNA:1.99±0.19 vs.1.00±0.11,P<0.001),cell viability decreased by approximately 28%(P<0.001),and lipid peroxides increased[(13.34±3.20)%vs.(5.92±2.14)%]The level of malondialdehyde(MDA)increased[(1.70±0.19)nmol/mgprot vs.(0.91±0.03)nmol/mgprot,P<0.001].The content of reduced glutathione(GSH)decreased[(6.64±1.64)μmol/mgprot vs.(16.84±2.45)μmol/mgprot,P<0.001].Compared with the control group,the number of migratory cells in the Erastin group was significantly reduced(283.20±52.78 vs.410.20±76.01,P=0.015),and the number of invasive cells was significantly reduced(119.60±36.46 vs.240.40±40.20,P=0.001).The results of western blotting showed that compared with the control group,the expression level of E-cadherin protein in the Erastin group was upregulated(1.12±0.10 vs.0.56±0.09,t=9.290,P<0.001).Vimentin(0.81±0.05 vs.1.10±0.10,t=5.605,P<0.001),N-cadherin(0.38±0.06 vs.0.72±0.11,t=6.010,P<0.001)and GPX4(0.15±0.06 vs.0.37±0.07,t=5.862,P<0.001)were downregulated.The lipid peroxidation activity of cells in the oe-ACSL4 group increased,the lipid peroxidation activity in the oe-ACSL4+Erastin group further increased,and the level of lipid peroxidation in the oe-ACSL4+Fer-1 group decreased,and the difference was statistically significant(P<0.001).Compared with the control group,the MDA level in the oe-ACSL4 group increased,while the protein expression levels of GSH and GPX4 decreased.Compared with the oe-ACSL4 group,the MDA level in the oe-ACSL4+Fer-1 group decreased,while the GSH content and GPX4 protein expression increased.Conclusion Low expression of ACSL4 is significantly associated with poor prognosis of ovarian cancer.From a mechanistic perspective,ACSL4 can inhibit the proliferation,migration and invasion of tumor cells by enhancing ferroptosis activity.
Keywords：ovarian canceracyl-CoA synthetase long-chain family member 4ferroptosislipid peroxidation
Publication Date：2025-12-28
Online Publishing Date：2026-01-12(First online date of this platform, not the publication date of the document)
Pages：7( 1145-1151 )