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目的检测肿瘤抑制基因p16INK4A在小鼠动情周期和早孕期子宫内膜中的表达,初步探讨p16INK4A在小鼠胚胎植入过程中的生物学作用。方法运用反转录-聚合酶链反应(RT-PCR)技术检测动情周期和早孕期小鼠子宫内膜p16INK4A mRNA的表达水平;用免疫组织化学和Western blotting方法检测p16INK4A蛋白的表达。结果RT-PCR显示,早孕期比动情周期p16INK4A mRNA的表达有显著增强。动情周期中动情期比其他3期表达明显增强;早孕期中,随妊娠天数的增加,p16INK4A mRNA的表达也逐渐增强,孕5d达峰值,之后又逐渐减弱。免疫组织化学和Western blotting的结果也都显示,p16INK4A蛋白在子宫内膜的表达及规律与RT-PCR的结果一致。结论p16INK4A在小鼠的胚泡植入过程中起重要作用。
Objective To detect the expression of tumor suppressor gene p16INK4A in mouse estrous cycle and early pregnancy endometrium, and to explore the biological role of p16INK4A in mouse embryo implantation. Methods Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of p16INK4A mRNA in estrous cycle and early pregnancy mice. The expression of p16INK4A protein was detected by immunohistochemistry and Western blotting. Results RT-PCR showed that the expression of p16INK4A mRNA in early pregnancy was significantly higher than that in estrous cycle. Estrus cycle, estrus than the other three significantly enhanced expression; early pregnancy, with the increase of the number of days of pregnancy, p16INK4A mRNA expression also gradually increased, reaching a peak on 5d, and then gradually weakened. Immunohistochemistry and Western blotting results also show that p16INK4A protein expression in the endometrium and the law and RT-PCR results. Conclusion p16INK4A plays an important role in blastocyst implantation in mice.