印记基因P57在小鼠乳腺发育和泌乳中的功能及其机制初步研究

发布时间：2018-08-30 14:15

【摘要】：印记基因是一类亲本来源特异性表达的基因,大部分印记基因在胚胎发育、胎盘形成以及个体生活过程中扮演着重要角色。P57是周期蛋白激酶抑制剂,是一个典型的抑癌基因,同时也是一个父本印记、母本表达的印记基因,其缺失会带来胚胎致死。P57免疫荧光染色显示其蛋白在乳腺组织中广泛表达,RT-PCR结果表明它在乳腺形态变化的4个时期中表达量呈下降趋势。为了研究P57在乳腺发育过程中是否起到了作用,本文在乳腺细胞中设计了两种shRNA的对P57进行了敲除,发现P57通过影响乳腺上皮细胞的干性来影响增殖,RT结果显示其干细胞信号通路的相关基因表达水平明显降低,然而周期检测发现其G1期的细胞比例下降。通过对临近LncRNA Kcnq1ot1进行敲除可以使P57的表达量显著上调,但对PRC2复合物成员Suz12敲除虽然可以降低Kcnq1ot1的表达,但P57的变化趋势却不稳定。在对P57进行敲除后,RT检测到另外15个印记基因的表达水平也发生显著变化,验证了乳腺细胞中印记基因网络的存在。
[Abstract]:Imprinted genes are a class of genes specifically expressed by their parents. Most imprinted genes play an important role in embryonic development, placenta formation and individual life. P57 is a cyclin kinase inhibitor and a typical tumor suppressor gene. It's also a paternal imprint, the imprinted gene expressed by the mother. The results of RT-PCR showed that the expression of P57 protein decreased in the four stages of mammary gland morphological changes. In order to study whether P57 plays a role in breast development, we designed two kinds of shRNA to knock out P57 in mammary gland cells. It was found that P57 affected the proliferation of mammary epithelial cells by affecting the dryness of mammary epithelial cells. The results showed that the expression level of genes related to stem cell signaling pathway was significantly decreased, but the proportion of cells in G1 phase was decreased by cycle detection. The expression of P57 could be upregulated significantly by knockout of adjacent LncRNA Kcnq1ot1, but Suz12 knockout of PRC2 complex member could decrease the expression of Kcnq1ot1, but the change trend of P57 was not stable. The expression level of the other 15 imprinted genes was also significantly changed by RT after p57 knockout, which confirmed the existence of imprinted gene network in breast cells.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q78

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