β1，4-半乳糖基转移酶I与细胞间黏附分子1在着床调控中的相关性探讨

发布时间：2018-01-26 20:39

  本文关键词： ICAM-1 β1 4-GalT-I 着床调控　出处：《大连医科大学》2010年硕士论文　论文类型：学位论文

【摘要】： 目的：探讨着床前子宫内膜细胞β1,4-GalT-I的表达及调控β1,4-GalT-I对细胞间黏附分子1(ICAM-1)表达的影响,分析β1,4-GalT-I与ICAM一1在胚胎着床过程中的相关性。 β1,4-半乳糖基转移酶I(β1,4-GalT-I)分布于高尔基体和细胞膜两种亚细胞结构上,位于高尔基体上的β1,4-GalT-I可行使蛋白质的糖基化修饰功能；位于细胞膜上的β1,4-GalT-I则作为一种细胞黏附分子,参与细胞黏附、精卵识别、神经轴突的生长、肿瘤细胞迁移等过程。 细胞间黏附分子1(intercellular adhesion molecule-1,ICAM-1)既是一类重要的黏附分子,也是一种跨膜糖蛋白,它在早期胚泡发育、子宫内膜容受性的建立及胚泡着床过程中有重要作用。 方法：利用人源的子宫内膜癌细胞(RL95-2)模拟着床前子宫内膜,人源的绒毛癌细胞(JAR)模拟胚泡来组成体外着床模型,将β1,4-GalT-I基因过表达质粒(HA-GalT-I)以及空载体质粒(pCDNA3.0)转染RL95-2细胞使其过表达β1,4-半乳糖基转移酶Ⅰ,通过细胞荧光检测确定转染效果,采用RT-PCR、Western-blot等方法检测调控β1,4-GalT-I表达对ICAM-1表达的影响,检测JAR细胞在RL95-2细胞上的黏附率判断二者在胚胎着床调控中的相关性。 结果：转染β1,4-GaT-I过表达质粒HA-GalT-I后60小时,RL95-2细胞中有绿色荧光蛋白的表达,RT-PCR及Western-blot分析结果显示：与正常培养组、空质粒转染组相比,过表达HA-GalT-I组β14-GaT-I的表达量增高,ICAM-I的基因表达和蛋白合成也随之增加;过表达HA-GalT-I后JAR细胞在RL95-2细胞上的黏附率(78.5%)也明显高于正常培养组(65.4%)和空质粒转染组(62.5%),(p0.05),说明上调β1,4-GalT-I会促进ICAM-I表达及提高胚泡着床率。 结论： (1)调控β1,4-GalT-I表达可影响ICAM-1的表达； (2)β1,4-GalT-I可能通过调节ICAM-1的表达参与着床调控。
[Abstract]:Objective: to investigate the expression of 尾 _ 1N _ 4-GalT-I and its effect on the expression of intercellular adhesion molecule 1 (ICAM-1) in endometrial cells before implantation, and to analyze the effect of 尾 _ 1 on the expression of intercellular adhesion molecule 1 (ICAM-1). The correlation between 4-GalT-I and ICAM-1 during embryo implantation. 尾 _ 1 ~ (4-galactosyltransferase) I (尾 _ 1N _ 4-GalT-I) distributes in Golgi apparatus and cell membrane, and is located on 尾 _ 1 on Golgi body. 4-GalT-I can perform the glycosylation modification function of protein. As a cell adhesion molecule, 尾 _ 1N _ 4-GalT-I, located on the cell membrane, is involved in cell adhesion, sperm-egg recognition, axon growth, tumor cell migration and so on. Intercellular adhesion molecule-1 (ICAM-1) is an important adhesion molecule. It is also a transmembrane glycoprotein, which plays an important role in the development of early blastocyst, the establishment of endometrial receptivity and the implantation of blastocyst. Methods: human endometrial cancer cells (RL95-2) were used to simulate preimplantation endometrium, and human chorionic carcinoma cells (JARs) were used to simulate blastocyst to form an in vitro implantation model. The overexpression plasmid HA-GalT-I of 4-GalT-I and pCDNA3.0) were transfected into RL95-2 cells to overexpression 尾 1. The transfection effect of 4-galactosyltransferase 鈪，

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