ll-trans retinoic acid (ATRA) can reverse the malignant behaviors of hepatocellular carcinoma (HCC) cells

thereby exerting anti-HCC effect; however

the underlying mechanism is yet to be understood. This study aimed to demonstrate that ATRA is vital to ferroptosis in HCC. Ferroptosis-related genes exhibit different expression in patients with HCC compared to that in healthy individuals. A total of 20 amino acid products were detected in HepG2 cells

the expression level of 5 was decreased after ATRA treatment. ATRA improved the levels of lipid ROS

MDA

and NAPD

+

/NADPH

and reduced the mt-DNA copy number and changed the structure of mitochondria

in HepG2 and Hep3B cells. We found the expression of genes positively correlated with ferroptosis to increase and those negatively correlated to decrease with ATRA treatment. Inhibition of ferroptosis by Ferrostatin-1 reversed ATRA-inhibited proliferation of HCC cells

along with cell migration and invasion. GSH synthesis was blocked by ATRA

accompanied by decreased cystine content and increased glutamate content

and downregulation of the expression of GSH synthesis-related genes. Our findings suggested that ATRA inhibited the malignancy of HCC cells by improving ferroptosis

and that inhibition of GSH synthesis contributed to ATRA-induced ferroptosis.