肿瘤转移抑制因子CD82/KAI1对植入前小鼠胚胎体外发育的影响

目的:研究CD82/KAI1mRNA及蛋白质在小鼠胚胎中的表达规律及其对小鼠胚胎体外发育的影响。方法:①应用RT-PCR及免疫荧光技术观察CD82/KAI1mRNA及蛋白质在小鼠不同发育时期胚胎中的表达规律。②在不同CD82/KAI1抗体浓度的培养液中,培养小鼠8-细胞胚胎,观察囊胚发育率、孵化率和胚胎细胞数的变化。结果:①CD82/KAI1mRNA在小鼠不同时期胚胎中均有表达,于8-细胞期及桑葚胚表达较丰富,CD82/KAI1蛋白表达于各期胚胎细胞的胞膜和胞浆,在桑葚胚期CD82蛋白还强表达于胚胎细胞的胞核。②一定浓度的CD82/KAI1抗体可明显抑制胚胎的发育速度(1∶800,P<0.05),降低囊胚的形成率(1∶400,P<0.05)和孵出率(1∶800,P<0.05),减少囊胚细胞数(1∶800,P<0.05)。结论:CD82/KAI1在小鼠胚胎的发育过程中可能发挥重要作用。

Effect of Tumor Metastasis Inhibitory Factor CD82/KAI1 Gene on in vitro Development of Pre-implantation Embryo in Mouse

Objective: To identify expression of tumor metastasis suppressor gene CD82/KAI1 mRNA and protein in mouse embryos, and its effect on in vitro development of mouse embryos. Methods: 1) Expressions of tumor metastasis suppressor gene CD82/KAI1 mRNA and protein were detected by RT-PCR and immunofluorescence. 2) 8-cell embryos were cultured at various concentrations of CD82/KAI1 antibody, and the percentage of embryo cleavage, blastocyst hatching and total cell number of each blastocyst were counted respectively. Results: 1) Both CD82/KAI1 mRNA and protein were expressed in all stages of embryo development. CD82/KAI1 protein was detected in cellular membrane and cytoplasm. At morula stage, CD82/KAI1 protein was mainly expressed in cell nucleus. 2) In vitro culture, CD82 antibody significantly inhibited embryo developing from 8-cell to blastocyst stage (1∶400, P<0.05), and dramatically decreased rate of blastocyst hatching (1∶800, P <0.05) and total cell number of each blastocyst in a dose-dependent manner. Conclusion: CD82/KAI1 might play an important role in embryo development.