论文部分内容阅读
为探讨DNA结合蛋白果蝇Eph激酶(Drosophila Eph kinase,DEK)对人子宫内膜基质蜕膜化的调节作用和途径,该研究采用qPCR(Real-time quantitative polymerase chain reaction)、免疫组化(immunohistochemical)和蛋白印迹(Western blot)分别检测人子宫内膜增生期、分泌期和蜕膜组织中DEK基因和蛋白的表达;利用siRNA抑制基质细胞和蜕膜细胞的DEK,再用细胞流式技术、细胞免疫荧光、qPCR、细胞碱性磷酸酶脂显色和Western blot检测DEK沉默后细胞的变化。结果显示,蜕膜组织中DEK mRNA表达水平低于增生期和分泌期(P<0.05),蜕膜组织中DEK蛋白表达水平高于增生期和分泌期(P<0.05);抑制基质细胞DEK会使细胞增殖和分化能力降低,从而抑制基质细胞蜕膜化;抑制蜕膜细胞DEK可使细胞凋亡增加,DNA损伤情况加剧,导致蜕膜细胞的维持和发展受到影响。综上,该研究初步证实,DEK可能通过调控细胞蜕膜化而参与胚胎着床过程,其可能途径与其通过调控细胞增殖、分化、凋亡和核损伤有关。
To investigate the regulatory effect and mechanism of Drosophila Eph kinase (DEK), a DNA-binding protein, on human endometrial stromal degeneration, we used qPCR (real-time quantitative polymerase chain reaction), immunohistochemistry ) And Western blot were used to detect the expression of DEK gene and protein in the proliferative, secretory and decidual tissues of human endometrium respectively. SiRNA was used to inhibit the DEK of stromal cells and decidual cells, Cell immunofluorescence, qPCR, cell alkaline phosphatase, and Western blot were used to detect the changes of cells after DEK silencing. The results showed that the expression level of DEK mRNA in decidual tissue was lower than that in proliferative phase and secretory phase (P <0.05), and the level of DEK protein in decidual tissue was higher than that in proliferative phase and secretory phase (P <0.05) So that the proliferation and differentiation of cells to reduce, thereby inhibiting stromal cell decidualization; inhibit decidual DEK cells can increase apoptosis, DNA damage increased, leading to the maintenance and development of decidual cells affected. In summary, the study initially confirmed that DEK may participate in embryo implantation by regulating cell decidualization, which may be related to its regulation of cell proliferation, differentiation, apoptosis and nuclear injury.