INMAP overexpression inhibits cell proliferation, induces genomic instability and functions through p53/p21 pathways

• Published in: PloS one Vol. 10; no. 1; p. e0115704
• Main Authors: Zhu, Yan, Lei, Yan, Du, Baochen, Zheng, Yanbo, Lu, Xiangfeng, Tan, Tan, Kang, Jingting, Sun, Le, Liang, Qianjin
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 30.01.2015; Public Library of Science (PLoS)
• Subjects: Agar; Analysis; Angiogenesis; Animals; Apoptosis; Bcl-2 protein; Biochemical markers; Biology; Cell Cycle Proteins - biosynthesis; Cell Cycle Proteins - genetics; Cell growth; Cell proliferation; Cell Proliferation - genetics; Chromosomes; Colorectal cancer; Cyclin-dependent kinase inhibitor p21; Cyclin-Dependent Kinase Inhibitor p21 - biosynthesis; Dehydrogenases; Deoxyribonucleic acid; DNA; DNA damage; Education; Experiments; Gene expression; Gene Expression Regulation; Genomic Instability; GTP-binding protein; HeLa Cells; Histones - biosynthesis; Humans; Hypoxia; Immunofluorescence; Immunoglobulins; Isoenzymes; L-Lactate dehydrogenase; Laboratories; Lactate dehydrogenase; Lactic acid; Life sciences; Medical prognosis; Mice; Micronuclei; Mitosis; Monomolecular films; Nuclear Proteins - biosynthesis; Nuclear Proteins - genetics; p53 Protein; Proliferating cell nuclear antigen; Proteins; Signal Transduction - genetics; Spindle Apparatus - genetics; Stability; Tumor proteins; Tumor Suppressor Protein p53 - biosynthesis; Tumors; Xenograft Model Antitumor Assays; Xenografts
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0115704

INMAP is a spindle protein that plays essential role for mitosis, by ensuring spindle and centromere integrality. The aim of this study was to investigate the relevant functions of INMAP for genomic stability and its functional pathway. We overexpressed INMAP in HeLa cells, resulting in growth inhibition in monolayer cell cultures, anchorage-independent growth in soft agar and xenograft growth in nude mice. In this system caused micronuclei (MNi) formation, chromosome distortion and γH2AX expression upregulation, suggesting DNA damage induction and genomic stability impairment. As a tumour biochemical marker, lactate dehydrogenase (LDH) isoenzymes were detected to evaluate cell metabolic activity, the results confirming that total activity of LDH, as well as that of its LDH5 isoform, is significantly decreased in INMAP-overexpressing HeLa cells. The levels of p53 and p21 were upregulated, and however, that of PCNA and Bcl-2, downregulated. Indirect immunofluorescence (IIF) and coimmunoprecipitation (CoIP) analyses revealed the interaction between INMAP and p21. These results suggest that INMAP might function through p53/p21 pathways.