【摘要】 目的探讨CD82对着床窗口期小鼠子宫内膜上皮细胞内整合素αV、β3、E-cadherin以及β-catenin蛋白表达的影响。方法将构建的CD82腺病毒转染原代培养的小鼠子宫内膜上皮细胞。检测妊娠小鼠子宫内膜上皮细胞转染CD82腺病毒后,细胞内整合素αV、β3、E-cadherin和β-catenin的表达变化情况。结果提取的上皮细胞纯度为(93.2±0.6)%。构建的CD82腺病毒转染效率达到(92.0±4.5)%,转染原代培养的小鼠子宫内膜上皮细胞24 h后,RT-PCR检测发现CD82基因表达明显升高。转染48 h后,Western blot检测CD82蛋白水平明显升高。免疫细胞化学检测妊娠第4天的小鼠子宫内膜上皮细胞转染CD82腺病毒后,整合素αV、β3以及β-catenin的表达较未转染组均有明显上升(P<0.05),但E-cadherin的表达量无明显变化(P>0.05)。结论胚胎植入前,CD82可能影响小鼠子宫内膜上皮细胞内整合素αV、β3和β-catenin的蛋白表达。更多还原

【Abstract】 Objective Uterine epithelial cells were isolated from pregnant mice and cultured in vitro,and examined the effect of CD82 on the expression of integrin αV,β3,E-cadherin and β-catenin in the cells. Methods The uterine epithelial cells were primarily isolated from pregnant mouse uterus. The recombinant adenovirus containing mouse CD82gene which had been constructed in our lab infected the uterine epithelial cells. Immunocytochemistry was used to detect the protein expressions of integrin αV,β3,E-cadherin and β-catenin in the uterine epithelial cells,which were infected with CD82 adenovirus or not. Results 1. The purity of primary cultured cells was( 93. 2 ± 0. 6) %. 2. The transfection efficiency of CD82 recombinant adenovirus was( 92 ± 4. 5) %. The adenoviral particles carrying CD82 gene indeed expressed CD82 gene and protein in the primary uterine epithelial cells after 24 hours or 48 hours. 3. The uterine epithelial cells of pregnant mice on d4 expressed integrin αV,β3,E-cadherin and β-catenin. 4. In contrast to the control group,when CD82 adenovirus infected cells,the uterine epithelial cells of pregnant mice on d4 increased the expression of integrinαV,β3 and β-catenin protein,had no significant changes of E-cadherin. Conclusions CD82 may have effect on the expression of integrin αV,β3 and β-catenin in mouse uterine epithelial cells before implantation.更多还原