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Published on: February 6, 2018
Knockdown of polycomb-group RING finger 6 modulates mouse male germ cell differentiation in vitro
1Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, Shanghai, China.
Background:
Polycomb-group RING finger 6 (PCGF6), one of six PCGF homologs, is the core component of the PRC1 complex that plays critical roles in epigenetic transcriptional silencing in higher eukaryotes. However, the biological functions of PCGF6 are unknown.
Method:
qRT-PCR and Western blot were used to detect the expression profile of PCGF6 in testes. Fluorescent immunohistochemistry was used to examine the cellular localization of PCGF6 protein in testes. Cell proliferation was tested by performing CCK-8 and EdU incorporation assay. Cell cycle and haploid cell population analysis was determined by flow cytometry using propidium iodide DNA staining. Co-immunoprecipitation experiment was conducted using PCGF6 antibody to obtain interacting protein of PCGF6. Luciferase reporter assays were performed to examine the promoter activity of PCGF6 in cells overexpressing OVOL1 and PLZF.
Results:
PCGF6 was expressed predominantly in meiotic and post-meiotic male germ cells, could negatively regulate the proliferation of GC-2 spd cells, an immortalized mouse spermatogenic cell line, and could modulate the differentiation of GC-2 spd cells in vitro. PCGF6 could indirectly interact with HSPA2, a key factor that is essential for male meiosis, and OVOL1 and PLZF, two key transcription factors that are involved in spermatogenesis, could positively and negatively modulate Pcgf6 promoter activity, respectively. PCGF1, BMI1, and PCGF5 are also highly expressed in mouse testes like PCGF6.
Conclusion:
PCGF6 may play important roles in male germ cell development.
