Disorder of G2-M Checkpoint Control in Aniline-Induced Cell Proliferation in Rat Spleen

• Published in: PloS one Vol. 10; no. 7; p. e0131457
• Main Authors: Wang, Jianling, Wang, Gangduo, Khan, M Firoze
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 20.07.2015; Public Library of Science (PLoS)
• Subjects: Aberration; Aniline; Aniline Compounds - toxicity; Animals; Biocompatibility; Cancer therapies; Carcinogenesis - drug effects; CDC2 Protein Kinase; Cell cycle; Cell growth; Cell proliferation; Cell Proliferation - drug effects; Control; Cyclin A - genetics; Cyclin A - metabolism; Cyclin B - genetics; Cyclin B - metabolism; Cyclin-dependent kinase; Cyclin-Dependent Kinase Inhibitor p21 - metabolism; Cyclin-dependent kinase inhibitor p27; Cyclin-Dependent Kinase Inhibitor p27 - metabolism; Cyclin-dependent kinases; Cyclin-Dependent Kinases - metabolism; Cyclins; Deoxyribonucleic acid; Deregulation; DNA; DNA damage; Down-Regulation - drug effects; Drinking water; Epigenetic inheritance; Exposure; Fibrosis; G1 phase; G2 phase; G2 Phase Cell Cycle Checkpoints - drug effects; Gene expression; Humans; Hyperplasia; Inhibitors; Iron; Kinases; M Phase Cell Cycle Checkpoints - drug effects; Male; Mammals; MicroRNA; MicroRNAs; MicroRNAs - genetics; Molecular modelling; Oxidative stress; Pathology; Phosphorylation; Proteins; Rats; Rats, Sprague-Dawley; Regulatory proteins; RNA, Messenger - genetics; RNA, Messenger - metabolism; Rodents; Spleen; Spleen - cytology; Spleen - drug effects; Spleen - metabolism; Splenocytes; Splenomegaly; Studies; Toxicity; Tumor markers; Tumors
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0131457

Aniline, a toxic aromatic amine, is known to cause hemopoietic toxicity both in humans and animals. Aniline exposure also leads to toxic response in spleen which is characterized by splenomegaly, hyperplasia, fibrosis and the eventual formation of tumors on chronic in vivo exposure. Previously, we have shown that aniline exposure leads to iron overload, oxidative DNA damage, and increased cell proliferation, which could eventually contribute to a tumorigenic response in the spleen. Despite our demonstration that cell proliferation was associated with deregulation of G1 phase cyclins and increased expression of G1 phase cyclin-dependent kinases (CDKs), molecular mechanisms, especially the regulation of G2 phase and contribution of epigenetic mechanisms in aniline-induced splenic cellular proliferation remain largely unclear. This study therefore, mainly focused on the regulation of G2 phase in an animal model preceding a tumorigenic response. Male Sprague-Dawley rats were given aniline (0.5 mmol/kg/day) in drinking water or drinking water only (controls) for 30 days, and expression of G2 phase cyclins, CDK1, CDK inhibitors and miRNAs were measured in the spleen. Aniline treatment resulted in significant increases in cell cycle regulatory proteins, including cyclins A, B and CDK1, particularly phosphor-CDK1, and decreases in CDK inhibitors p21 and p27, which could promote the splenocytes to go through G2/M transition. Our data also showed upregulation of tumor markers Trx-1 and Ref-1 in rats treated with aniline. More importantly, we observed lower expression of miRNAs including Let-7a, miR-15b, miR24, miR-100 and miR-125, and greater expression of CDK inhibitor regulatory miRNAs such as miR-181a, miR-221 and miR-222 in the spleens of aniline-treated animals. Our findings suggest that significant increases in the expression of cyclins, CDK1 and aberrant regulation of miRNAs could lead to an accelerated G2/M transition of the splenocytes, and potentially to a tumorigenic response on chronic aniline exposure.