MiRNA-133b promotes the proliferation of human Sertoli cells through targeting GLI3.

Chencheng Yao,Zheng Chen,Qingqing Yuan,Min Sun,Minghui Niu,Hong Wang,Yun Liu,Jingmei Hou,Zheng Li,Liping Wen,Zuping He

doi:10.18632/oncotarget.6876

Abstract

Sertoli cells play critical roles in regulating spermatogenesis and they can be reprogrammed to the cells of other lineages, highlighting that they have significant applications in reproductive and regenerative medicine. The fate determinations of Sertoli cells are regulated precisely by epigenetic factors. However, the expression, roles, and targets of microRNA (miRNA) in human Sertoli cells remain unknown. Here we have for the first time revealed that 174 miRNAs were distinctly expressed in human Sertoli cells between Sertoli-cell-only syndrome (SCOS) patients and obstructive azoospermia (OA) patients with normal spermatogenesis using miRNA microarrays and real time PCR, suggesting that these miRNAs may be associated with the pathogenesis of SCOS. MiR-133b is upregulated in Sertoli cells of SCOS patients compared to OA patients. Proliferation assays with miRNA mimics and inhibitors showed that miR-133b enhanced the proliferation of human Sertoli cells. Moreover, we demonstrated that GLI3 was a direct target of miR-133b and the expression of Cyclin B1 and Cyclin D1 was enhanced by miR-133b mimics but decreased by its inhibitors. Gene silencing of GLI3 using RNA inference stimulated the growth of human Sertoli cells. Collectively, miR-133b promoted the proliferation of human Sertoli cells by targeting GLI3. This study thus sheds novel insights into epigenetic regulation of human Sertoli cells and the etiology of azoospermia and offers new targets for treating male infertility

Highlights

Non-obstructive azoospermia (NOA), which is defined as the absence of sperm in the ejaculation resulted from the testicular failure, affects about 10% of infertile men, and the incidence of NOA has been found in almost 60% of the azoospermia patients [1]
We have recently revealed that stem cell factor (SCF), bone morphogenic protein 4 (BMP4), and Glial cell line-derived neurotrophic factor (GDNF) are differentially expressed in human Sertoli cells between NOA patients and OA patients with normal spermatogenesis [13] and that BMP4 promotes the proliferation of human Sertoli cells through the Smad1/5 and ID2/3 pathway [14], which provides novel insights into genetic etiology of NOA azoospermia
After removing male germ cells, human Sertoli cells were cultured in Dulbecco modified Eagle medium (DMEM)/F12 supplemented with 10% fetal bovine serum (FBS) for one day

Summary

Introduction

Non-obstructive azoospermia (NOA), which is defined as the absence of sperm in the ejaculation resulted from the testicular failure, affects about 10% of infertile men, and the incidence of NOA has been found in almost 60% of the azoospermia patients [1]. Spermatogenesis is an intricate process, including the self-renew and differentiation of spermatogonial stem cells (SSCs), the meiosis of spermatocytes, and the spermiogenesis of haploid spermatids. These three phases are under the intimate modulation of testicular microenvironment or the ‘niche’. We have recently revealed that SCF, BMP4, and GDNF are differentially expressed in human Sertoli cells between NOA patients and OA patients with normal spermatogenesis [13] and that BMP4 promotes the proliferation of human Sertoli cells through the Smad1/5 and ID2/3 pathway [14], which provides novel insights into genetic etiology of NOA azoospermia. The epigenetic regulation of human Sertoli cells needs to be clarified

Methods

Results

Discussion

Conclusion