RESUMO
Mouse blastocysts were exposed to doses of 0, 1 and 10 mumol/L retinoic acid (RA) for 24 h and the cytotoxic effect of RA on the mouse blastocysts in vitro was observed. FITC-labeled terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL-FITC) assay was employed to stain apoptotic cells and immunohistochemical S-P staining method was used to detect the expression of Fas protein in mouse blastocysts in vitro. The results showed that RA could induce apoptosis and increase the expression of Fas proteins of trophectoderm (TE) and inner cell mass (ICM) cells in blastocysts. Compared with the findings for the control blastocysts, exposure to RA (10 mumol/L) resulted in a more significant apoptosis and higher expression level of Fas proteins (P<0.01). It was concluded that RA could induce apoptosis, which may result in a significant reduction in the average number of total cells and the trophectoderm/inner cell mass in blastocysts and an increased expression of Fas protein, suggesting that RA had a cytotoxic effect on the growth and development of early embryos in mice.
Assuntos
Receptor fas/biossíntese , Apoptose/efeitos dos fármacos , Blastocisto/citologia , Blastocisto/metabolismo , Técnicas de Cultura de Células/métodos , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Marcação In Situ das Extremidades Cortadas , RNA Mensageiro/metabolismo , Tretinoína/farmacologiaRESUMO
To investigate the expressions of Fas and Fas ligand (FasL) in human placenta, we studied the expressions of Fas and FasL in placenta with RT-PCR, immunoblotting and immunostaining. We observed amplified products of Fas and FasL transcripts, the band of Fas (52 kDa) and multiple bands of FasL (42-52 kDa) in pla-centa. Fas and FasL localized mainly on fetal vessels and on syncytiotrophoblasts respectively. The differential distribution of Fas and FasL in human placenta may reflect intrinsic expressions of them by trophoblasts during differentiation. The increased expression of Fas in trophoblasts may promote apoptosis of placenta in pathologic condition such as preeclampsia.
Assuntos
Humanos , Receptor fas/biossíntese , Proteína Ligante Fas , Expressão Gênica , Perfilação da Expressão Gênica , Glicosilação , Immunoblotting/métodos , Glicoproteínas de Membrana/biossíntese , Placenta/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trofoblastos/citologiaRESUMO
Flow cytometric immunofluorescent analysis was used to assess Fas antigen [CD95] expression in blasts obtained from bone marrow of 24 patients with acute myeloid leukemia. The percentage of positive cells in each sample was highly variable. Fas antigen expression did nor correlate with age Hb% concentration, while blood cell count, platelet count, blast% in peripheral blood, blast% in bone marrow, CD34 expression or BC12 protein. Low expression of Fas was associated with a low complete remission rate after induction chemotherapy [69.2% in cases with <20% positive cells versus 90.9% in cases with >/20% positive cells, P <0.01]. The main cause for not achieving remission was resistant diseases. The result of this study suggested that the quantitation of Fas expression can be predictive of treatment outcome in acute myeloid leukemia