Genistein up-regulates miR-20a to disrupt spermatogenesis via targeting Limk1

// Hao Gu 1, 2, 3, * , Wei Wu 1, 2, 4, * , Beilei Yuan 1, 2 , Qiuqin Tang 5 , Dan Guo 1, 2 , Yiqiu Chen 1, 2 , Yankai Xia 1, 2 , Lingqing Hu 4 , Daozhen Chen 4 , Jiahao Sha 6 and Xinru Wang 1, 2 1 State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China 2 Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China 3 Department of Central Laboratory, Huai’an First People’s Hospital, Nanjing Medical University, Huai’an 223002, China 4 State Key Laboratory of Reproductive Medicine, Wuxi Maternal and Child Health Care Hospital Affiliated to Nanjing Medical University, Wuxi 214002, China 5 State Key Laboratory of Reproductive Medicine, Department of Obstetrics, Obstetrics and Gynecology Hospital Affiliated to Nanjing Medical University, Nanjing 210004, China 6 State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 211166, China * These authors contributed equally to the study and they should be regarded as joint first authors Correspondence to: Wei Wu, email: wwu@njmu.edu.cn Qiuqin Tang, email: t19871004@sina.com Keywords: genistein, miR-17-92 cluster, miR-20a, Limk1, spermatogenesis Received: September 26, 2016      Accepted: April 16, 2017      Published: May 05, 2017 ABSTRACT Genistein (GEN) is one of the isoflavones that has effect on male reproduction. However, the underlying mechanism remains unknown. miRNAs are a type of small non-coding RNAs that play important roles in spermatogenesis. We measured the GEN levels and miR-17-92 cluster expression in infertile subjects and found that miR-17-92 might be involved in GEN induced abnormal spermatogenesis. To clarify, we fed adult ICR mice with different doses of GEN (0, 0.5, 5, 50 and 250 mg/kg/day) for 35 days to study the underlying mechanism. We found that sperm average path velocity, straight-line velocity and eurvilinear velocity of the mice orally with GEN at 5mg/kg/day were significantly decreased, the expression levels of miR-17 and miR-20a in mice testis were higher in corresponding group. We also found miR-20a was the only miRNA that differentially expressed both in human and mice. By applying bioinformatics methods, Limk1 was predicted to be the target gene of miR-20a that is involved in spermatogenesis. Limk1 were significantly decreased in the corresponding group. Dual-luciferase report assay also proved that miR-20a could directly target Limk1. These results implied that Limk1 might be the target gene of miR-20a that is involved in GEN induced abnormal spermatogenesis.