Abstract: OBJECTIVE To investigate whether WBSCR22 was involved in ferroptosis. METHODS The siRNA targeting human WBSCR22 gene(siWBSCR22) was designed and synthesized, which was transient transfected to knockdown WBSCR22 mRNA. The level of WBSCR22 mRNA and protein were measured by RT-qPCR, Western blotting, respectively. The human colorectal cancer cell lines HCT116, RKO, lung cancer cell lines H460, and hepatoma cell line SMMC7721 were treated with the ferroptosis inducer Erastin, RSL3 alone or in combination with antioxidant NAC, necrosis inhibitor NSA, apoptosis inhibitor ZVAD-FMK, autophagy inhibitor 3-MA, and cell death rate was measured by CCK8. In addition, the cellular lipid peroxidation was measured by MDA assay and the cellular concentration of Fe²⁺ was measured by Fe assay. RESULTS Transient transfection of siWBSCR22 significantly reduced the mRNA and protein levels of WBSCR22. WBSCR22 knockdown significantly promoted the ferroptosis induced by Erastin and RSL3 in HCT116, RKO, H460, and SMMC7721 cells; the addition of NAC significantly reduced ferroptosis, while NSA, ZVAD-FMK, and 3-MA had no effect on ferroptosis. Erastin or RSL3 treatment significantly increased lipid peroxidation in cancer cells, which led to ferroptosis. However, Erastin or RSL3 did not alter the Fe²⁺ concentration in cells. CONCLUSION WBSCR22 gene regulates ferroptosis in cancer cells.