| ObjectiveThe analysis of genetic expression profile in spontaneous abortion deciduas with applied gene array found that signal transducers and activators of transcription 1 (STAT1) gene was one of the significant up-regulation expression genes. STAT1 protein that is regarded as a key link of JAK/STAT1 signal transduction pathway plays an important role in adjusting transcription and expression processes of relevant gene. A number of studies have identified that lots of cytokines, such as IFN, IL-2, IL-4, IL-6, CNTF and growth factors, such as EGF, PDGF, CSF utilize this signal transduction pathway to induce cell proliferation, differentiation or Apoptosis. But there is no research to explore effect of STAT1 in early pregnant deciduas. On the basis of our previous research, this study aims to identify STAT1 expression in deciduas from normal pregnancy and spontaneous abortion at gene level and protein level respectively, in order to explore the role of STAT1 in implantation and spontaneous abortion.Methods1.Group of decidua from normal pregnancy(1) Collecting decidua of artificial abortion during the 6th week to 11th week of early normal pregnancy respectively.(2) Freezing decidua at -80℃and extracting total RNA of each decidua, then detecting STAT1 mRNA expression of the decidua samples by RT-PCR.(3) Observing STAT1 mRNA expression directly by in-situ hybridization. (4) Oobserving STAT1 expression of the decidua samples directly by immunohistochemistry and immunofluorescence.(5) Freezeing decidua at -80℃and extracting total protein of each decidua, and then detecting STAT1 protein expression of the decidua samples by western blot .2.Group of decidua from spontaneous abortion(1) Collecting decidua from spontaneous abortion respectively, including missed abortion, inevitable abortion and threatened abortion (during the 6th week to 11th week of early pregnancy) .(2) The decidua were analyzed by RT-PCR, immunohistochemistry, western blot, in-situ hybridization and immunofluorescence. Results 1. Group of decidua from normal pregnancy (1) The results indicated that STAT1 mRNA expression in deciduas showed a gradual descent trend with days past by RT-PCR.(2) The findings of in-situ hybridization showed that STAT1 mRNA expressed in decidua cell's endochylema. STAT1 mRNA expression was the most strong in the 6th week of pregnancy and then showed gradual descent trend with days past, but the most weak in the 11th week of pregnancy.(3) The findings from western blot were similar with what have been found by RT-PCR, that STAT1 protein expression showed gradual descent trend with days past.(4) The results from immunohistochemistry and immunofluorescence had identified STAT1 expression in decidua cell's endochylema. STAT1 expression was the most strong in the 6th week of pregnancy and then showed gradual descent trend with days past., but the most weak in the 11th week of pregnancy.2. Group of decidua from spontaneous abortion(1) The findings from RT-PCR showed that there is no significant difference on STAT1 mRNA expression in decidua from three kind of samples of spontaneous abortion, including missed abortion,inevitable abortion and threatened abortion, but their expression were stronger than that of decidua from normal pregnancy.(2) The results from in-situ hybridization showed that STAT1 mRNA widely distributed in endochylema and cell nucleus and there was no significant difference among three kind of decidua from spontaneous abortion, but their expression were significant stronger than that in deciduas from normal pregnancy.(3) The findings from western blot showed that there is no difference on STAT1 protein expression in three kind of decidua from spontaneous abortion, but their expressions were significant stronger than that in decidua from normal pregnancy.(4) The results from immunohistochemistry and immunofluorescence showed that STAT1 protein widely distributed in endochylema and cell nucleus of three kind of decidua from spontaneous abortion, and there was no significant difference among three kind of deciduas from spontaneous abortion,but their color were significant deeper than that of deciduas from normal pregnancy.ConclusionSemi-quantization RT-PCR, immunohistochemistry, in-situ hybridization, Western Blot and immunofluorescence were applied to detect STAT1 expression in deciduas from spontaneous abortion and normal pregnancy. It showed that STAT1 expression showed gradual descent trend with days past in decidus from normal pregnancy during the 6th week to the 11th week, but STAT1 expression was significant stronger in decidus from spontaneous abortion than that in decidus from normal pregnancy. These results suggested that STAT1 might play an important role in the implantation. Abnormal increasing of STAT1 expression may affect implantation through some mechanisms, and then further induce abortion. |
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