title: Nucleolin down-regulation is involved in ADP-induced cell cycle arrest in S phase and cell apoptosis in vascular endothelial cells.
abstract: High concentration of extracellular ADP has been reported to induce cell apoptosis, but the molecular mechanisms remain not fully elucidated. In this study, we found by serendipity that ADP treatment of human umbilical vein endothelial cells (HUVEC) and human aortic endothelial cells (HAEC) down-regulated the protein level of nucleolin in a dose- and time-dependent manner. ADP treatment did not decrease the transcript level of nucloelin, suggesting that ADP might induce nucleolin protein degradation. HUVEC and HAEC expressed ADP receptor P2Y13 receptor, but did not express P2Y1 or P2Y12 receptors. However, P2Y1, 12, 13 receptor antagonists MRS2179, PSB0739, MRS2211 did not inhibit ADP-induced down-regulation of nucleolin. Moreover, MRS2211 itself down-regulated nucleolin protein level. In addition, 2-MeSADP, an agonist for P2Y1, 12 and 13 receptors, did not down-regulate nucleolin protein. These results suggested that ADP-induced nucleolin down-regulation was not due to the activation of P2Y1, 12, or 13 receptors. We also found that ADP treatment induced cell cycle arrest in S phase, cell apoptosis and cell proliferation inhibition via nucleolin down-regulation. The over-expression of nucleolin by gene transfer partly reversed ADP-induced cell cycle arrest, cell apoptosis and cell proliferation inhibition. Furthermore, ADP sensitized HUVEC to cisplatin-induced cell death by the down-regulation of Bcl-2 expression. Taken together, we found, for the first time to our knowledge, a novel mechanism by which ADP regulates cell proliferation by induction of cell cycle arrest and cell apoptosis via targeting nucelolin.
authors.affiliation: Department of Internal Medicine, Hunan Armed Police Force's Hospital, Changsha, Hunan, China; Medical Research Center, Changsha Central Hospital, Changsha, Hunan, China.
authors.forename: Wenmeng
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Internal Medicine, Hunan Armed Police Force's Hospital, Changsha, Hunan, China.
authors.forename: Junqing
authors.initials: J
authors.lastname: Luo
authors.affiliation: Department of Internal Medicine, Hunan Armed Police Force's Hospital, Changsha, Hunan, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Xiang
authors.affiliation: Medical Research Center, Changsha Central Hospital, Changsha, Hunan, China.
authors.forename: Xueting
authors.initials: X
authors.lastname: Liu
authors.affiliation: Medical Research Center, Changsha Central Hospital, Changsha, Hunan, China.
authors.forename: Manli
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Medical Research Center, Changsha Central Hospital, Changsha, Hunan, China.
authors.forename: Lingjuan
authors.initials: L
authors.lastname: Liao
authors.affiliation: Medical Research Center, Changsha Central Hospital, Changsha, Hunan, China.
authors.forename: Jinyue
authors.initials: J
authors.lastname: Hu
mesh_terms: D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D001011:Aorta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D001391:Azo Compounds / Q000494:pharmacology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010750:Phosphoproteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D058913:Purinergic Agonists / Q000494:pharmacology; D058914:Purinergic Antagonists / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000378:metabolism; D058472:Receptors, Purinergic P2Y1 / Q000172:deficiency / Q000235:genetics; D058925:Receptors, Purinergic P2Y12 / Q000172:deficiency / Q000235:genetics; D059807:S Phase Cell Cycle Checkpoints / Q000187:drug effects* / Q000235:genetics; D015398:Signal Transduction; D013873:Thionucleotides / Q000494:pharmacology; D000097796:Nucleolin
chemical_list: D000244:Adenosine Diphosphate; D000970:Antineoplastic Agents; D001391:Azo Compounds; D002945:Cisplatin; D010750:Phosphoproteins; D019253:Proto-Oncogene Proteins c-bcl-2; D058913:Purinergic Agonists; D058914:Purinergic Antagonists; D011732:Pyridoxal Phosphate; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D058925:Receptors, Purinergic P2Y12; D013873:Thionucleotides; D000097796:Nucleolin; C000595707:BCL2 protein, human; C556691:MRS 2211; C548576:P2RY12 protein, human; C440036:P2RY13 protein, human; C002480:methylthio-ADP