microRNA-383 impairs phosphorylation of H2AX by targeting PNUTS and inducing cell cycle arrest in testicular embryonal carcinoma cells.

Abstract

Male germ cells with aberrant DNA damage are the weighted factor contributing to male infertility. Mounting evidence shows that DNA damage in male germ cells impairs spermatogenesis and lowers fecundity. MicroRNAs (miRNAs) regulating expression of multiple genes play a significant role in spermatogenesis. Our previous results have shown that microRNA-383 (miR-383) is one of the notable down-regulated microRNAs in the testes of sterile males with maturation arrest (MA) and is located predominantly in spermatogonia and primary spermatocytes. However, the role that miR-383 plays in DNA damage during spermatogenesis remains unknown. In this study, we found that miR-383 inhibited the focal formation and abundance of γH2AX, which is the major marker of sites of DNA damage, with or without ultraviolet irradiation and cisplatin in testicular embryonal carcinoma (NT-2) cells. In addition, NT-2 cells were remarkably sensitized to DNA damage reagent (cisplatin) by forcing expression of miR-383 and silencing expression of protein phosphatase 1, regulatory subunit 10 (PNUTS). By constructing Renilla luciferase reporters and co-transfecting miR-383 and reporters in NT-2 cells, we identified that PNUTS was a valid target of miR-383. Further results demonstrated that the repression of the phosphorylated form of H2AX by miR-383 was due to independent depletion of PNUTS and cell cycle arrest. In conclusion, we found a novel function of miR-383 in the DNA damage pathway. miR-383 impairs the phosphorylation of H2AX by targeting PNUTS and inducing cell cycle arrest independently, as well as sensitizing NT-2 cells to cisplatin.

• Authors: Helong Huang, Hui Tian, Zhengzheng Duan, Yunxia Cao, Xian-Sheng Zhang, Fei Sun
• Journal: Cellular signalling (Cell Signal) Vol. 26 Issue 5 Pg. 903-11 (May 2014) ISSN: 1873-3913 [Electronic] England
• PMID: 24462707 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2014 Elsevier Inc. All rights reserved.

Chemical References

• Antineoplastic Agents
• DNA-Binding Proteins
• H2AX protein, human
• Histones
• MIRN383 microRNA, human
• MicroRNAs
• Nuclear Proteins
• PPP1R10 protein, human
• RNA-Binding Proteins
• Protein Phosphatase 1
• Cisplatin

Topics

• Adult
• Antineoplastic Agents (toxicity)
• Cell Cycle Checkpoints (drug effects, genetics, radiation effects)
• Cell Line, Tumor
• Cisplatin (toxicity)
• DNA Damage (drug effects, radiation effects)
• DNA-Binding Proteins (antagonists & inhibitors, genetics, metabolism)
• Embryonal Carcinoma Stem Cells (cytology, metabolism)
• Histones (metabolism)
• Humans
• Male
• MicroRNAs (metabolism)
• Neoplasms, Germ Cell and Embryonal (metabolism, pathology)
• Nuclear Proteins (antagonists & inhibitors, genetics, metabolism)
• Phosphorylation (genetics)
• Protein Phosphatase 1 (genetics, metabolism)
• RNA-Binding Proteins (antagonists & inhibitors, genetics, metabolism)
• Testicular Neoplasms (metabolism, pathology)
• Ultraviolet Rays