Nucleolin down-regulation is involved in ADP-induced cell cycle arrest in S phase and cell apoptosis in vascular endothelial cells

• Published in: PloS one Vol. 9; no. 10; p. e110101
• Main Authors: Wang, Wenmeng, Luo, Junqing, Xiang, Fang, Liu, Xueting, Jiang, Manli, Liao, Lingjuan, Hu, Jinyue
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 07.10.2014; Public Library of Science (PLoS)
• Subjects: Adenosine diphosphate; Adenosine Diphosphate - analogs & derivatives; Adenosine Diphosphate - pharmacology; Antineoplastic Agents - pharmacology; Aorta; Aorta - cytology; Aorta - drug effects; Aorta - metabolism; Apoptosis; Apoptosis - drug effects; Azo Compounds - pharmacology; Bcl-2 protein; Biology and Life Sciences; Cancer; Cell adhesion & migration; Cell cycle; Cell death; Cell growth; Cell Line; Cell proliferation; Cell Proliferation - drug effects; Cisplatin; Cisplatin - pharmacology; Deoxyribonucleic acid; DNA; Dose-Response Relationship, Drug; Down-regulation; Endothelial cells; Endothelial Cells - cytology; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Endothelium; Gene expression; Gene Expression Regulation; Gene transfer; Human Umbilical Vein Endothelial Cells - cytology; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - metabolism; Humans; Inhibition; Internal medicine; Kinases; Leukemia; Medical research; Medicine and Health Sciences; Molecular modelling; Monocytes - cytology; Monocytes - drug effects; Monocytes - metabolism; Nucleolin; Overexpression; Phosphoproteins - antagonists & inhibitors; Phosphoproteins - genetics; Phosphoproteins - metabolism; Primary Cell Culture; Proteins; Proteolysis; Proto-Oncogene Proteins c-bcl-2 - antagonists & inhibitors; Proto-Oncogene Proteins c-bcl-2 - genetics; Proto-Oncogene Proteins c-bcl-2 - metabolism; Purinergic Agonists - pharmacology; Purinergic Antagonists - pharmacology; Pyridoxal Phosphate - analogs & derivatives; Pyridoxal Phosphate - pharmacology; Receptors; Receptors, Purinergic P2 - genetics; Receptors, Purinergic P2 - metabolism; Receptors, Purinergic P2Y1 - deficiency; Receptors, Purinergic P2Y1 - genetics; Receptors, Purinergic P2Y12 - deficiency; Receptors, Purinergic P2Y12 - genetics; RNA, Messenger - antagonists & inhibitors; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA-Binding Proteins - antagonists & inhibitors; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; S phase; S Phase Cell Cycle Checkpoints - drug effects; S Phase Cell Cycle Checkpoints - genetics; Signal Transduction; Thionucleotides - pharmacology; Transcription; Umbilical vein; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0110101

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.