关键词: 成人；睾丸；支持细胞； 增殖；β-catenin； c-myc

Abstract:

Objective   To investigate the effect and mechanism of Wnt / β-catenin signaling pathway on adult human Sertoli cell proliferation. Methods   Adult testicular tissues were digested by the two-step enzymatic method to extract and purify Sertoli cells, and the purified Sertoli cells were continuously passage cultured. The Sertoli cell-specific protein vimentin was detected by immunofluorescence to evaluate the purity of the cultured Sertoli cells. The cultured Sertoli cells were divided into three groups: the normal culture medium (Con group), the medium with Gsk-3β inhibitor SB216763（SB group）, and the medium with lithium chloride (LiCl group).  BrdU incorporation assay was performed to measure Sertoli cell proliferation and flow cytometry was used to access the cell cycle. Immunofluorescence, real-time fluorescent quantitative PCR and Western blotting were performed to investigate expressions of β-catenin and c-myc. C-myc expression was down regulated in Sertoli cells by small RNA interference technology. Results   BrdU incorporation assay and flow cytometry results revealed that SB216763 and the LiCl significantly promoted Sertoli cell proliferation. The nuclear translocation of β-catenin and c-myc, the mRNA level and protein expression of c-myc remarkably increased in SB group and the LiCl group. The proliferation stimulation effect of SB216763 on human Sertoli cells was significantly inhibited after c-myc siRNA transfection. Conclusion   Activation of the Wnt/β-catenin pathway could promote the proliferation of the adult testicular Sertoli cells via upregulation of c-myc expression.

Key words: Adult; Sertoli cells; Proliferation; β-catenin; C-myc