〔Abstract〕 Objective To explore the mechanism of Golgi phosphoprotein 3(GOLPH3) regulating lysosomal biogenesisviamTORC1 signaling.Methods GOLPH3 knock-out(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 compared.Results GOLPH3 KO suppressed mTORC1 activity significantly, decreased the cytoplasm level of pTFEB, increased the nuclear localization of active TFEB, and promoted the lysosome biogenesis.Conclusion Golgi protein GOLPH3 regulates lysosome biogenesis through mTORC1 signaling.