title: The anticoagulant effect of PGI2S and tPA in transgenic umbilical vein endothelial cells is linked to up-regulation of PKA and PKC.
abstract: The selection of vascular grafts for coronary artery bypass surgery is crucial for a positive outcome. This study aimed to establish a novel line of vascular endothelial cells with a potent anticoagulant effect. A lentiviral vector was used to stably transfect human umbilical vein endothelial cells (HUVECs) with PGI2S alone (HUVEC-PGI2S) or both PGI2S and tPA (HUVEC-PGI2S-tPA). Both HUVEC-PGI2S and HUVEC-PGI2S-tPA cells over-expressing PGI2S and tPA were compared to mock-transfected cells. The enzyme-linked immuno sorbent assay (ELISAs) demonstrated that the anticoagulation components, ATIII and PLG, were up-regulated and coagulation factor FVIII was down-regulated in both cell lines. QRT-PCR and western blotting demonstrated the vasodilation and platelet disaggregation proteins PKA, PKC, and PTGIR were up-regulated in both cell lines, but MAPK expression was not altered in either cell line. However, cell viability and colony formation assays and cell cycle analysis demonstrated that both cell lines had a lower rate of cell growth and induced G1 phase arrest. HUVEC-PGI2S and HUVEC-PGI2S-tPA cells have a potent anticoagulant effect and their use in vascular heterografts may decrease the risk of thrombosis.
authors.affiliation: Cardiovascular Surgery Department, Second People's Hospital of Guangdong Province, 1 Shi-liu Gang Road, Guangzhou 510317, China. JianhuaWang2014@gmail.com.
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Oncology in Southern China, Sun Yat-sen University Cancer Center, 651 Dongfeng Road East, Guangzhou 510060, China. yuanlj@sysucc.org.cn.
authors.forename: Lin-Jing
authors.initials: LJ
authors.lastname: Yuan
authors.affiliation: Cardiovascular Surgery Department, Second People's Hospital of Guangdong Province, 1 Shi-liu Gang Road, Guangzhou 510317, China. zmin908@sina.com.
authors.forename: Zhi-Min
authors.initials: ZM
authors.lastname: Zhong
authors.affiliation: Department of Chest, Sun Yat-sen University Cancer Center, 651 Dongfeng Road East, Guangzhou 510060, China. wenzhsh@sysucc.org.cn.
authors.forename: Zhe-Sheng
authors.initials: ZS
authors.lastname: Wen
authors.affiliation: Department of Chest, Second People's Hospital of Guangdong Province, 1 Shi-liu Gang Road, Guangzhou 510317, China. DoctorDengjianming@126.com.
authors.forename: Jian-Ming
authors.initials: JM
authors.lastname: Deng
authors.affiliation: Cardiovascular Surgery Department, Second People's Hospital of Guangdong Province, 1 Shi-liu Gang Road, Guangzhou 510317, China. shiliyingsly@sina.com.
authors.forename: Rong-Xin
authors.initials: RX
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Oncology in Southern China, Sun Yat-sen University Cancer Center, 651 Dongfeng Road East, Guangzhou 510060, China. zhengmin@sysucc.org.cn.
authors.forename: Min
authors.initials: M
authors.lastname: Zheng
mesh_terms: D000925:Anticoagulants / Q000494:pharmacology*; D000990:Antithrombin III / Q000378:metabolism; D002470:Cell Survival; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D011464:Epoprostenol / Q000235:genetics / Q000378:metabolism*; D005169:Factor VIII / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D044006:Receptors, Epoprostenol; D011982:Receptors, Prostaglandin / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D010959:Tissue Plasminogen Activator / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D000925:Anticoagulants; C113936:PTGIR protein, human; D044006:Receptors, Epoprostenol; D011982:Receptors, Prostaglandin; D000990:Antithrombin III; D005169:Factor VIII; D011464:Epoprostenol; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D010959:Tissue Plasminogen Activator