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目的:探讨MAD2、BUB1 mRNA及蛋白表达水平在人类胚胎染色体分离中的作用。方法:应用荧光原位杂交(fluorescence in situ hybridization,FISH)技术检测102例自然流产胚胎绒毛染色体数目,将自然流产胚胎分为染色体数目正常组和异常组,分别用RT-PCR和Western blotting检测胚胎绒毛组织中MAD2、BUB1 mRNA和蛋白质相对表达水平。结果:①102例自然流产胚胎中检出染色体数目异常50例(49.02%),其中多倍体9例、非整倍体36例、嵌合型5例。②异常组胚胎绒毛中MAD2 mRNA及蛋白相对表达水平分别为0.89±0.19和0.45±0.06,均明显高于正常组(0.80±0.21和0.42±0.08,P<0.05)。③异常组胚胎绒毛中BUB1 mRNA及蛋白相对表达水平分别为1.04±0.23和0.35±0.05,与正常组(0.98±0.25和0.33±0.07)比较,差异无统计学意义(P>0.05)。结论:胚胎染色体数目异常是导致自然流产的重要原因;MAD2在胚胎染色体数目异常组中高表达,延长了纺锤体组装检查点(SAC)的有丝分裂阻滞持续时间,在保证染色体正确分离、维持基因组稳定方面起重要作用。BUB1是SAC的上游基因,在延长有丝分裂阻滞中不起主要作用,但其正常表达对于SAC发挥功能必不可少。
Objective: To investigate the role of MAD2, BUB1 mRNA and protein expression in the separation of human embryos. Methods: Fluorescence in situ hybridization (FISH) was used to detect the number of chorionic villi in 102 spontaneous abortion embryos. The spontaneous abortions were divided into normal group and abnormal group. The embryos were detected by RT-PCR and Western blotting Relative expression levels of MAD2, BUB1 mRNA and protein in villous tissue. Results: ① The number of chromosomal abnormalities detected in 102 spontaneous abortions was 50 (49.02%), including 9 polyploidy, 36 aneuploidy, and 5 chimerism. ② The relative expression levels of MAD2 mRNA and protein in the abnormal group were 0.89 ± 0.19 and 0.45 ± 0.06, which were significantly higher than those in the normal group (0.80 ± 0.21 and 0.42 ± 0.08, P <0.05). ③ The relative expression levels of BUB1 mRNA and protein in abnormal group were 1.04 ± 0.23 and 0.35 ± 0.05, respectively. There was no significant difference between normal group (0.98 ± 0.25 and 0.33 ± 0.07) (P> 0.05). Conclusion: Abnormal number of embryos is an important cause of spontaneous abortion. MAD2 is overexpressed in the abnormal number of embryos, prolongs the duration of mitotic arrest at spindle assembly checkpoint (SAC), ensures correct chromosome segregation and maintains genomic stability Play an important role. BUB1, an upstream gene of SAC, plays a major role in prolonging mitotic arrest, but its normal expression is essential for the functioning of SACs.