You Xinguo, Wei Lu, Fan Shutong, Yang Weiwei, Liu Xiaoying, Wang Guohui, Man Yi, Pan Zhifang, Feng Weiguo
College of Bioscience and Technology, Weifang Medical University, Weifang, Shandong 261053, P.R. China.
Mol Med Rep. 2017 Aug;16(2):2101-2106. doi: 10.3892/mmr.2017.6797. Epub 2017 Jun 16.
Zinc finger protein 185 (ZNF185) belongs to the ZNF family and is involved in cell proliferation and differentiation. To the best of our knowledge, the association between ZNF185 and male reproduction is unknown. In the present study, the expression and localization of ZNF185 in mouse testis, as well as its role in testosterone secretion, cell cycle progression and apoptosis of mouse Leydig cells were investigated. The results of the immunofluorescence analysis indicated that ZNF185 was highly expressed in Leydig cells of the mouse testis, and primarily localized in the cytoplasm. The results of quantitative polymerase chain reaction and western blot analyses further validated that ZNF185 expression was significantly higher in Leydig cells and sperm compared with that in Sertoli cells. Subsequently, the expression pattern of ZNF185 in mouse testis was determined at different developmental stages. The results demonstrated that the expression of ZNF185 was highest in the testis of 10‑week‑old mice and lowest in 2‑week‑old mice. Furthermore, the role of ZNF185 in Leydig cells of the mouse testis was investigated. Different concentrations of luteinizing hormone (LH) were used to stimulate the Leydig cells and subsequently the expression of ZNF185, and testosterone concentration was detected. The results revealed that LH upregulated the expression of ZNF185 and testosterone secretion, and ZNF185 expression was significantly positively correlated with testosterone secretion. To further validate whether ZNF185 was involved in testosterone secretion, lentiviral‑mediated RNA interference was used to knock down ZNF185 expression in Leydig cells. The results demonstrated that ZNF185 expression and testosterone secretion of Leydig cells were decreased significantly. In addition, the results demonstrated that the knockdown of ZNF185 expression did not significantly affect cell cycle progression or apoptosis. Taken together, the results of the present study revealed that ZNF185 was highly expressed in Leydig cells of the testis and involved in the secretion of testosterone. These results have contributed to the elucidation of the mechanism underlying male reproduction and may provide a novel target for the treatment of infertility, and the development of a contraceptive vaccine.
锌指蛋白185(ZNF185)属于锌指蛋白家族,参与细胞增殖和分化。据我们所知,ZNF185与雄性生殖之间的关联尚不清楚。在本研究中,我们调查了ZNF185在小鼠睾丸中的表达和定位,以及它在小鼠睾丸间质细胞睾酮分泌、细胞周期进程和凋亡中的作用。免疫荧光分析结果表明,ZNF185在小鼠睾丸间质细胞中高表达,主要定位于细胞质。定量聚合酶链反应和蛋白质印迹分析结果进一步证实,与支持细胞相比,ZNF185在睾丸间质细胞和精子中的表达显著更高。随后,我们确定了ZNF185在小鼠睾丸不同发育阶段的表达模式。结果表明,ZNF185在10周龄小鼠睾丸中的表达最高,在2周龄小鼠睾丸中的表达最低。此外,我们研究了ZNF185在小鼠睾丸间质细胞中的作用。使用不同浓度的促黄体生成素(LH)刺激睾丸间质细胞,随后检测ZNF185的表达和睾酮浓度。结果显示,LH上调了ZNF185的表达和睾酮分泌,且ZNF185表达与睾酮分泌显著正相关。为了进一步验证ZNF185是否参与睾酮分泌,我们使用慢病毒介导的RNA干扰技术敲低睾丸间质细胞中ZNF185的表达。结果表明,睾丸间质细胞中ZNF185的表达和睾酮分泌显著降低。此外,结果表明敲低ZNF185的表达并未显著影响细胞周期进程或凋亡。综上所述,本研究结果表明,ZNF185在睾丸间质细胞中高表达,并参与睾酮分泌。这些结果有助于阐明雄性生殖背后的机制,并可能为不孕症的治疗和避孕疫苗的开发提供新的靶点。