过表达CD82对植入期小鼠子宫内膜整合素αV、β3、E-cadherin和β-catenin表达的影响  

Effect of CD82 on the expression of integrin αV,β3,E-cadherin and β-catenin in uterine epithelial cells in pregnant mice

作　　者：尉晓蔚[1] 谭冬梅[2] 张倩[2] 何通川[3] 谭毅[2] 

机构地区：[1]大连大学附属中山医院,大连116001 [2]重庆医科大学实验动物中心,重庆400016 [3]美国芝加哥大学分子肿瘤实验室

出　　处：《中国实验动物学报》2014年第3期57-61,I0011-I0012,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(30470654);重庆市自然科学基金(2009BB5409);教育部博士点科研基金(20095503110006)

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。Objective Uterine epithelial cells were isolated from pregnant mice and cultured in vitro , and exam-ined the effect of CD82 on the expression of integrin αV,β3, E-cadherin and β-catenin in the cells.Methods The uter-ine epithelial cells were primarily isolated from pregnant mouse uterus .The recombinant adenovirus containing mouse CD 82 gene 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 CD 82 gene indeed ex-pressed 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 d 4 increased the expression of integrinαV,β3 and β-catenin protein, had no significant changes of E-cadherin.Conclusions CD82 may have effect on the ex-pression of integrin αV,β3 and β-catenin in mouse uterine epithelial cells before implantation .

关 键 词：CD82基因 重组腺病毒 子宫内膜上皮细胞 原代培养 孕小鼠 

分 类 号：Q95-33[生物学—动物学] Q492