Knockdown of Polycomb-Group RING Finger 6 Modulates Mouse Male Germ Cell Differentiation in Vitro
| Autor: | Yi Lin, Ji Wu, Lin He, Jian Wang, Jin Sun |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Male
Physiology Cellular differentiation Kruppel-Like Transcription Factors Biology PCGF6 lcsh:Physiology Cell Line lcsh:Biochemistry chemistry.chemical_compound Mice Testis medicine Animals Humans Germ cell development HSP70 Heat-Shock Proteins Promyelocytic Leukemia Zinc Finger Protein lcsh:QD415-436 Propidium iodide RNA Messenger RNA Small Interfering Promoter Regions Genetic Spermatogenesis Transcription factor Cellular localization Cell Proliferation Polycomb Repressive Complex 1 lcsh:QP1-981 Cell growth Cell Differentiation Cell cycle Molecular biology Spermatids DNA-Binding Proteins Mice Inbred C57BL medicine.anatomical_structure Germ Cells HEK293 Cells chemistry BMI1 Knockdown Germ cell Transcription Factors |
| Zdroj: | Cellular Physiology and Biochemistry, Vol 35, Iss 1, Pp 339-352 (2015) |
| ISSN: | 1421-9778 1015-8987 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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