肿瘤转移抑制因子CD82/KAI1在假孕小鼠子宫的表达及其对小鼠胚胎着床的影响

目的:研究CD82/KAI1mRNA及蛋白在假孕d1-8小鼠子宫中的表达规律,以及CD82/KAI1抗体对小鼠胚胎体外发育及着床的影响。方法:用RT-PCR及免疫组织化学技术观察CD82/KAI1mRNA及蛋白在假孕小鼠子宫的表达规律。将8-细胞小鼠胚胎培养于含不同浓度CD82/KAI1抗体的培养液中,观察囊胚形成及脱透明带的情况。妊娠d4小鼠宫角注射CD82/KAI1抗体,于妊娠d8观察小鼠胚胎植入数量。结果:①CD82/KAI1mRNA在假孕d1-8小鼠子宫中均有表达,于d4、d5表达最丰富。CD82/KAI1蛋白在假孕d1-8的子宫基质细胞及腺上皮细胞均有表达,假孕d1-4腔上皮细胞无表达,假孕d5子宫腔上皮细胞出现弱阳性表达,从d6开始子宫腔上皮细胞表达逐渐增强。②一定浓度的CD82/KAI1抗体可明显抑制囊胚的形成率(1∶400,P<0.05)和脱带率(1∶800,P<0.05)。③宫角注射一定量的CD82抗体可以显著提高小鼠胚胎的着床数(8.35±0.17vs4.52±0.24,P<0.05)。结论:CD82/KAI1在妊娠小鼠子宫的表达是非胚胎依赖性的。CD82/KAI1在小鼠胚胎发育和着床过程中发挥重要作用。

Spatiotemporal Expression of Tumor Metastasis Inhibitory Factor CD82/KAI1 Gene in Pseudopregnancy and Effect on Embryo Implantation in Mice

Objective:To identify mRNA and protein expression of tumor metastasis suppressor gene CD82/KAI1 in pseudopregnant mice and the effect of CD82/KAI1 polyclonal antibody on embryo development in vitro and implantation.Methods:Protein and mRNA expression of tumor metastasis suppressor gene CD82/KAI1 were detected by immunohistochemistry and RT-PCR.8-cell embryos were cultured in medium containing CD82/KAI1 antibody at various concentrations,then formation and hatching of blastocysts were observed.On d 8 the number of implantation sites was examined in uterine one horn,which was injected with CD82/KAI1 antibody on d 4.Results:(1)CD82/KAI1 mRNA was expressed during d 1-8 of pseudopregnancy,and it was dramatically abundant on d 4 and d 5.CD82/KAI1 protein was expressed mainly in stromal cells and glandular epithelial cells during d 1-8 of pseudopregnancy,however,no expression was detected in luminal epithelial cells during d 1-4 of pseudopregnancy.On d 5 weakly positive expression was observed,and the level of expression was gradually increased in luminal epithelial cells from d 6.(2)CD82 ployclonal antibody significantly inhibited formation of blastocysts(1:400,P<0.05)and prevented hatching of blastocysts from zona pellucida(1:800,P<0.05)in a dose-dependent manner.(3)The number of implanted embryos markedly increased in uterine one horn injected with CD82 antibody on d 4 of pregnancy(8.35±0.17 vs 4.52 ±0.24,P<0.05).Conclusion:Expression of CD82/KAI1 in uterus was independent of existence of embryos in pregnant mice.CD82/KAI1 might play an important role in embryo devel-opment and implantation in mice.