CD63基因在小鼠围着床期子宫中的表达和调节

发布时间：2018-01-11 17:02

本文关键词：CD63基因在小鼠围着床期子宫中的表达和调节　出处：《厦门大学》2009年硕士论文　论文类型：学位论文

【摘要】： 胚胎着床是一个复杂的过程,需要活化的胚泡与进入接受态的子宫内膜之间建立联系。在啮齿类动物,着床过程启动后,子宫内膜基质细胞发生广泛的蜕膜化,为接下来的胚胎及胎盘发育做准备。胚胎着床和子宫蜕膜化受到很多因素调控,但具体的分子机理仍不清楚。我们实验室利用基因表达系列分析(SAGE)技术发现,与非着床位点相比,CD63(CD63 antigen)基因在小鼠的着床位点显著上调。 CD63属于四次跨膜蛋白超家族,在人黑色素瘤等多种肿瘤发展中起重要作用,被称为肿瘤转移抑制因子。本实验利用原位杂交、实时定量RT-PCR、Westernblot和荧光素酶活性分析等方法,研究了CD63基因在围着床期小鼠子宫中的表达与调节。CD63的mRNA和蛋白水平在妊娠第5天子宫的着床位点均显著上调,在随后第6-8天的蜕膜区,均可检测到明显的CD63 mRNA表达信号。在假孕和延迟着床的子宫中,检测不到CD63基因的表达,而在延迟激活子宫的着床位点,CD63基因的表达特异性增强,其表达模式与正常妊娠第5天着床位点处类似。CD63 mRNA在小鼠妊娠第6-8天的蜕膜区表达,同时在人工诱导蜕膜化的小鼠子宫中也能检测到CD63基因的杂交信号,实时定量RT-PCR也验证了此结果,提示CD63基因可能在蜕膜化过程发挥作用。在切除卵巢的小鼠中,类固醇激素对CD63在子宫内的表达没有影响。体外诱导蜕膜化中,CD63 mRNA表达持续上调。CD63基因启动子上存在转录因子C/EBPβ反应元件,将含有CD63基因启动子序列的报告载体转染体外培养的子宫内膜基质细胞和NIH-3T3细胞后,通过荧光素酶活性分析,表明C/EBPβ能够显著增加CD63基因的转录活性。总之,CD63基因在小鼠围着床期子宫中的表达与胚胎着床过程密切相关,并受活性胚泡的诱导。CD63基因在小鼠子宫基质细胞体外蜕膜化过程中表达上调。此外,转录因子C/EBPβ上调CD63基因的转录活性。
[Abstract]:Embryo implantation is a complex process, which needs to establish a connection between the activated blastocyst and the recipient endometrium. In rodents, after implantation, the endometrial stromal cells undergo extensive decidualization. Prepare for subsequent embryo and placental development. Embryo implantation and uterine decidualization are regulated by many factors. But the specific molecular mechanism is still unclear. Our laboratory using gene expression series analysis of SAGE-based technology found that compared with non-implantation sites. CD63(CD63 antigenin gene was significantly up-regulated in mouse implantation site. CD63 belongs to the four-time transmembrane protein superfamily, and plays an important role in the development of human melanoma and other tumors, known as tumor metastasis suppressor. In this experiment, in situ hybridization, real-time quantitative RT-PCR. Methods of Westernblot and luciferase activity analysis. To study the expression and regulation of mRNA and protein of CD63 gene in the uterus of surrounding mice, the implantation sites of mRNA and CD63 were significantly up-regulated on the 5th day of pregnancy. The expression of CD63 mRNA was detected in decidua from the 6th to 8th day, and the expression of CD63 gene was not detected in pseudopregnancy and delayed implantation uterus. The expression of CD63 gene was specifically enhanced at the locus loci of delayed activation of uterus. The expression pattern of CD63 mRNA was similar to that at the implantation site on the 5th day of gestation in mouse decidua on the 6th and 8th day of gestation. At the same time, the hybridization signal of CD63 gene could also be detected in the mouse uterus induced by artificial decidualization, and the results were verified by real-time quantitative RT-PCR. These results suggest that CD63 gene may play a role in decidualization. Steroid hormones have no effect on the expression of CD63 in uterus in ovariectomized mice. The expression of CD63 mRNA continued to up-regulate. There were transcription factor C / EBP 尾 response elements on the promoter of CD63 gene. The reporter vector containing promoter sequence of CD63 gene was transfected into endometrial stromal cells and NIH-3T3 cells in vitro, and luciferase activity was analyzed. The results showed that C / EBP 尾 could significantly increase the transcriptional activity of CD63 gene. In conclusion, the expression of CD63 gene in mouse uterus is closely related to the implantation process. The expression of CD63 gene was up-regulated in mouse uterine stromal cells during decidualization in vitro, and the transcription factor C / EBP 尾 upregulated the transcriptional activity of CD63 gene.
【学位授予单位】：厦门大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R321;Q492

【共引文献】