Abstract: Abstract: Objective To establish a stable GFP-LC3-expressed human acute monocytic leukemia cell line (THP-1). Methods The lentivirus plasmid system (pCDH-CMV-GFP-LC3-EF1α-puro) was constructed and transfected into 293T cells with transfection reagent. THP-1 cells were infected with lentivirus and then screened by puromycin. The GFP-LC3 protein expression in THP-1 cells was analyzed by Western blot assay and flow cytometry. Starvation and rapamycin induced autophagy were detected by confocal microscope and Western blot assay. Results The THP-1 cell line with stable expressing GFP-LC3 protein showed visible green fluorescence under inverted fluorescence microscope, as demonstrated by Western blot assay and flow cytometry. Moreover, starvation and rapamycin both could induce the formation of autophagosomes, which included LC3 aggregates and LC3 modification. Conclusion The THP-1 cell line with stable expression of GFP-LC3 protein is constructed efficiently by the lentiviral plasmid system, which provides a cell model for autophagy detection in monocyte-macrophage.

Key words: autophagy, GFP-LC3, THP-1 cell, stable cell line, rapamycin