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本研究旨在检测肿瘤抑制基因p16INK4a(inhibitor of cyclin-dependent kinase4a)在早孕小鼠子宫内膜中的表达规律,探讨p16INK4a在小鼠胚胎着床过程中的作用。采用荧光定量PCR(FQ-PCR)和免疫组织化学方法分别检测未孕小鼠及孕小鼠第2、3、4、5、7天子宫内膜p16INK4a mRNA和蛋白的表达;子宫角注射p16INK4a抗体观察胚泡着床数。FQ-PCR结果显示孕小鼠子宫内膜组织p16INK4a mRNA的表达高于未孕小鼠,且随着妊娠天数的增加呈现表达逐渐增强的趋势,到妊娠第5天达到最高,后渐降。免疫组织化学分析显示p16INK4a蛋白在子宫内膜的表达规律与mRNA结果一致。子宫角注射p16INK4a抗体后胚泡着床数明显减少。以上结果提示,p16INK4a在妊娠早期子宫内膜持续表达,可能参与胚泡着床。
The purpose of this study was to investigate the expression of p16INK4a in the endometrium of early pregnant mice and to investigate the role of p16INK4a in the implantation of mouse embryos. The expression of p16INK4a mRNA and protein in endometrium was detected by fluorescence quantitative PCR (FQ-PCR) and immunohistochemical method on the 2nd, 3rd, 4th, 5th and 7th day respectively. The uterus was injected with p16INK4a antibody Observe the number of blastocyst implantation. The results of FQ-PCR showed that the expression of p16INK4a mRNA in endometrium of pregnant mice was higher than that of non-pregnant mice. The expression of p16INK4a increased gradually with the increase of gestational days and reached its peak on the 5th day of pregnancy and then decreased gradually. Immunohistochemical analysis showed that the expression of p16INK4a protein in the endometrium was consistent with the mRNA. Uterine injection p16INK4a antibody blastocyst significantly reduced the number of implantation. The above results suggest that p16INK4a is expressed continuously in the early stage of pregnancy and may be involved in blastocyst implantation.