RESUMO
Spermatogenesis requires both germ cells and testicular somatic cells, which are also involved in testicular development and male fertility. Sertoli cells are the only somatic cells in the seminiferous tubules and play very important roles in normal spermatogenesis. Abnormality of Sertoli cells in proliferation and adhesion may induce aberrant spermatogenesis and eventually cause infertility. Recently, various studies have demonstrated that miRNA are involved in the regulation of Sertoli cell proliferation and adhesion. Additionally, miRNA expression could be affected by hormone, endocrine interferon, and nutrition. In this review, we summarize miRNAs related to Sertoli cell proliferation and adhesion, which will be helpful for finding and identifying more miRNAs from Sertoli cells. The review will also provide theoretical basis for the pathogenesis of infertility associated with Sertoli cells.
Assuntos
Proliferação de Células , MicroRNAs/metabolismo , Células de Sertoli/citologia , Animais , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Masculino , MicroRNAs/genética , Túbulos Seminíferos/citologia , Túbulos Seminíferos/metabolismo , Células de Sertoli/metabolismo , EspermatogêneseRESUMO
OBJECTIVE: To construct a lentiviral expression vector of the PIAS-NY gene, and establish a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY. METHODS: PIAS-NY was synthesized, amplified by PCR and cloned into the lentiviral vector expression plasmid pGC-FU. After digestion and sequencing, pGC-FU-PIAS-NY, pHelper 1.0 and pHelper 2.0 were co-transfected into 293T cells. Then the lentiviral particles were used to transfect the mouse spermatocyte-derived cells. The expression of the PIAS-NY protein was detected by Western blot. RESULTS: We successfully constructed the lentiviral expression vector pGC-FU-PIAS-NY and established a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY. CONCLUSION: The construction of the lentiviral expression vector pGC-FU-PIAS-NY and the obtainment of stably transfected mouse spermatocyte-derived cells have paved the way for further studies on the roles of the PIAS-NY gene in spermatogenesis.
Assuntos
Linhagem Celular , Proteínas Inibidoras de STAT Ativados/genética , Espermatócitos/citologia , Transfecção , Animais , Vetores Genéticos , Lentivirus/genética , Masculino , Camundongos , PlasmídeosRESUMO
AIM: To prepare the mouse polyclonal antibody against human BRDT-NY prokaryotic protein and analyze the expression of BRDT-NY protein in digestive tract tumors. METHODS: The N-terminal amino acids of BRDT-NY protein was amplified by PCR and cloned into the expression vector pET28a(+). The recombinant plasmid pET28a+-BRDT-NY was transformed into E.coli BL21 and induced to express the recombinant protein with IPTG. We immunized BALB/c mice with the purified BRDT-NY protein for preparing the specific polyclonal antibody. The titer of the antibody was analyzed by ELISA. The expression of BRDT-NY protein in digestive tract tumors was detected by immunohistochemistry. RESULTS: Recombinant human BRDT-NY protein was expressed in BL21 and purified successfully by Ni-affinity chromatography. Two months after the mice were immunized with the purified BRDT-NY protein, we obtained the specific polyclonal antibody of high titer 1:100 000. Immunohistochemical analysis revealed that BRDT-NY protein was highly expressed in digestive tract tumors. CONCLUSION: The successful preparation of mouse polyclonal antibody against BRDT-NY lays a foundation for the research on the role of BRDT-NY protein in the pathology of human digestive tract tumors.
