title: Toxoplasma gondii ROP18: potential to manipulate host cell mitochondrial apoptosis.
abstract: Toxoplasma gondii is an obligate intracellular parasite that may manipulate host cell mitochondrial apoptosis pathways. In our experiment, 293T cells were transfected with the p3×FLAG-CMV-Myc-ROP18 vector and expressed the ROP18-Myc fusion protein. Cell apoptosis was induced by 0.5 μg/mL actinomycin D (ActD) and was detected by Annexin V-FITC/PI assay. The cell mitochondrial membrane potential was determined by JC-1. Cytochrome c (Cyto-c) from mitochondria and the cytoplasm was measured by Western blot. The Bcl-2 and Bax coding gene expression levels were detected by real-time PCR. We found, in vitro, that T. gondii ROP18 significantly suppressed 293T cell apoptosis induced by ActD and maintained mitochondrial membrane potential and integrity, thereby preventing the release of Cyto-c from mitochondria into the cytoplasm. The ratio of Bcl-2/Bax in ROP18-overexpressing cells was significantly higher than that of the negative control. Therefore, we speculate that ROP18 could suppress host cell apoptosis via the mitochondrial apoptosis pathway in vitro.
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, WHO Collaborating Center for Tropical Diseases, National Center of International Research on Tropical Diseases, Shanghai, 200025, China.|School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Wu
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Yunhui
authors.initials: Y
authors.lastname: Li
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Danhua
authors.initials: D
authors.lastname: Su
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Fu
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Liangping
authors.initials: L
authors.lastname: Gu
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Xugan
authors.initials: X
authors.lastname: Jiang
authors.affiliation: School of Medicine, Jiangsu University, Zhenjiang, 212013, China. chensxia@163.com.
authors.forename: Shengxia
authors.initials: S
authors.lastname: Chen
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, WHO Collaborating Center for Tropical Diseases, National Center of International Research on Tropical Diseases, Shanghai, 200025, China. caojp@yahoo.com.
authors.forename: Jianping
authors.initials: J
authors.lastname: Cao
mesh_terms: D017304:Annexin A5; D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D002460:Cell Line; D045304:Cytochromes c / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D057809:HEK293 Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000502:physiology*; D008928:Mitochondria / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D015800:Protozoan Proteins; D060888:Real-Time Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D017304:Annexin A5; C492760:FITC-annexin A5; C489427:MYC protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D016271:Proto-Oncogene Proteins c-myc; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D051028:bcl-2-Associated X Protein; D003609:Dactinomycin; D045304:Cytochromes c; D017346:Protein Serine-Threonine Kinases; C556793:ROP18 protein, Toxoplasma gondii; D016650:Fluorescein-5-isothiocyanate
keywords: Apoptosis; Mitochondrial apoptosis pathway; ROP18; Toxoplasma gondii