Objective : To explore the role of Golgi protein ACBD3 knockdown in ferroptosis. Methods : ACBD3 knockdown via shRNA , combined with treatment of ferroptosis inducer RSL3 , cell viability , iron levels , reactive oxygen species (ROS) levels , and lipid peroxide levels in different treatment groups were measured to comprehensively analyze the role of ACBD3 in ferroptosis. Results : Compared to control cells , the levels of ROS increased in ACBD3 knockdown cells. The total iron content was significantly reduced (P < 0. 001) , while the Fe2 + concentration in ACBD3 knockdown cells increased compared to control cells (P < 0. 001) . In the presence of ferroptosis inducer RSL3 , compared to control cells , cell viability significantly decreased ( P < 0. 001) , and lipid peroxide greatly increased in ACBD3 knockdown cells ( P < 0. 001) . Conclusion Golgi protein ACBD3 knockdown in mammalian cells changes iron homeostasis and promotes ferroptosis.

Objective : To explore the mechanism of Golgi phosphoprotein 3 (GOLPH3) regulating lysosomal biogenesis via mTORC1 signaling. Methods : GOLPH3 knockout (GOLPH3 KO) stable cell line was constructed by CRISPR Cas9. mTORC1 activity and the levels of TFEB and p ⁃TFEB in the control cells and GOLPH3 KO cells were compared through Western blot. Further, lysosome⁃associated membrane protein LAMP1 was labelled by the means of immunofluorescence and the number of lysosomes in the control cells and GOLPH3 KO cells was com⁃pared. Results : GOLPH3 KO suppressed mTORC1 activity significantly , decreased the cytoplasm level of p ⁃TFEB , increased the nuclear localization of active TFEB , and promoted the lysosome biogenesis. Conclusion Golgi protein GOLPH3 regulates lysosome biogenesis through mTORC1 signaling.