MiR-18a，miR-145和miR-495在小鼠胰腺发育过程中抑制Ptf1a的表达
发布时间：2018-10-15 06:47
【摘要】： 胰腺是一个重要的内外分泌混合腺,胰腺内分泌部中的β细胞缺失或发生功能障碍就会导致糖尿病,而外分泌细胞异常增殖会导致胰腺癌症的发生。因而研究胰腺器官发生发展的机制对于胰腺疾病的治疗具有重要意义。Ptfla(bHLH转录因子)是胰腺特化与外分泌分化所必需的转录因子。最近研究发现,Ptfla调控胰腺细胞的分化存在剂量依赖性:低水平的Ptfla促进内分泌分化,高水平Ptfla抑制内分泌分化促进外分泌分化。然而,胰腺不同发育命运细胞中Ptfla的表达调节机制还不清楚。 MicroRNAs(miRNAs)是一类22nt左右的非编码小RNA,它们在进化过程中高度保守,通过切割靶mRNA或抑制靶mRNA的翻译来调节基因的表达。一些研究已经表明,miRNAs在胰腺发育及功能调节中起重要作用,但目前还不知道哪些miRNAs在发育的胰腺上皮中调控Ptfla的表达。 为了解决这一问题,我们首先结合两个预测算法的结果,预测得到4个可能调控Ptfla表达的miRNAs。随后,我们通过双荧光素酶报告系统证明其中3个miRNAs(miR-18,miR-145和miR-495)确实对Ptfla的表达起到调控作用。通过qRT-PCR试验发现,在小鼠胰腺发育过程中miR-18a和Ptfla mRNA的表达模式成负相关;而免疫荧光染色试验显示miR-145/miR495与Ptfla蛋白的表达模式成负相关。最后,为了进一步了解小鼠胰腺发育过程中Ptfla相关的分子机制,我们结合以往文献数据与本文研究结果,绘制了小鼠胰腺发育中Ptfla相关的调控通路。 综上所述,本研究证明miR-18a,miR-145和miR-495可抑制Ptfla的表达,并参与到小鼠胰腺发育中Ptfla相关的调控网络中。
[Abstract]:The pancreas is an important mixture of internal and external secretory glands. The absence or dysfunction of 尾 cells in the endocrine region of the pancreas will lead to diabetes mellitus, while abnormal proliferation of exocrine cells will lead to the occurrence of pancreatic cancer. Therefore, it is important to study the mechanism of pancreatic organogenesis and development for the treatment of pancreatic diseases. Ptfla (bHLH transcription factors are essential transcription factors for pancreatic specialization and exocrine differentiation. Recently, it has been found that Ptfla regulates pancreatic cell differentiation in a dose-dependent manner: low level of Ptfla promotes endocrine differentiation, high level of Ptfla inhibits endocrine differentiation and promotes exocrine differentiation. However, the regulatory mechanism of Ptfla expression in cells with different developmental fate of pancreas is not clear that. MicroRNAs (miRNAs) is a kind of non-coding small RNA, around 22nt, which is highly conserved in evolution. Gene expression is regulated by cutting target mRNA or inhibiting target mRNA translation. Some studies have shown that miRNAs plays an important role in pancreatic development and function regulation, but it is not known which miRNAs regulates the expression of Ptfla in the developing pancreatic epithelium. In order to solve this problem, we first combine the results of two prediction algorithms to predict four miRNAs. that may regulate the expression of Ptfla. Subsequently, we demonstrated that three of the miRNAs (miR-18,miR-145 and miR-495) did regulate the expression of Ptfla by double luciferase reporting system. The expression pattern of miR-18a and Ptfla mRNA was negatively correlated with the expression pattern of miR-145/miR495 and Ptfla protein during the development of mouse pancreas by qRT-PCR assay, while the expression pattern of Ptfla protein was negatively correlated with the expression pattern of miR-145/miR495 by immunofluorescence staining. Finally, in order to further understand the molecular mechanism related to Ptfla during the development of mouse pancreas, we draw the regulatory pathway of Ptfla related to the development of mouse pancreas based on previous literature data and the results of this study. In conclusion, this study suggests that miR-18a,miR-145 and miR-495 can inhibit the expression of Ptfla and participate in the regulatory network related to Ptfla in the development of mouse pancreas.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R346
本文编号：2271707
[Abstract]:The pancreas is an important mixture of internal and external secretory glands. The absence or dysfunction of 尾 cells in the endocrine region of the pancreas will lead to diabetes mellitus, while abnormal proliferation of exocrine cells will lead to the occurrence of pancreatic cancer. Therefore, it is important to study the mechanism of pancreatic organogenesis and development for the treatment of pancreatic diseases. Ptfla (bHLH transcription factors are essential transcription factors for pancreatic specialization and exocrine differentiation. Recently, it has been found that Ptfla regulates pancreatic cell differentiation in a dose-dependent manner: low level of Ptfla promotes endocrine differentiation, high level of Ptfla inhibits endocrine differentiation and promotes exocrine differentiation. However, the regulatory mechanism of Ptfla expression in cells with different developmental fate of pancreas is not clear that. MicroRNAs (miRNAs) is a kind of non-coding small RNA, around 22nt, which is highly conserved in evolution. Gene expression is regulated by cutting target mRNA or inhibiting target mRNA translation. Some studies have shown that miRNAs plays an important role in pancreatic development and function regulation, but it is not known which miRNAs regulates the expression of Ptfla in the developing pancreatic epithelium. In order to solve this problem, we first combine the results of two prediction algorithms to predict four miRNAs. that may regulate the expression of Ptfla. Subsequently, we demonstrated that three of the miRNAs (miR-18,miR-145 and miR-495) did regulate the expression of Ptfla by double luciferase reporting system. The expression pattern of miR-18a and Ptfla mRNA was negatively correlated with the expression pattern of miR-145/miR495 and Ptfla protein during the development of mouse pancreas by qRT-PCR assay, while the expression pattern of Ptfla protein was negatively correlated with the expression pattern of miR-145/miR495 by immunofluorescence staining. Finally, in order to further understand the molecular mechanism related to Ptfla during the development of mouse pancreas, we draw the regulatory pathway of Ptfla related to the development of mouse pancreas based on previous literature data and the results of this study. In conclusion, this study suggests that miR-18a,miR-145 and miR-495 can inhibit the expression of Ptfla and participate in the regulatory network related to Ptfla in the development of mouse pancreas.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R346
【引证文献】
相关硕士学位论文 前2条
1 李丹;成体小鼠胰腺单细胞的离体分离、分析及培养[D];东北林业大学;2011年
2 杨艳坤;MicroRNA-18a在胰腺祖细胞和腺泡细胞中对转录因子Ptf1a的调控研究[D];东北林业大学;2012年
，本文编号：2271707
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