miR-92a-3p对前脂肪细胞增殖和分化的影响

发布时间：2018-02-11 16:59

本文关键词： miR-92a-3p 3T3-L1前脂肪细胞成脂分化增殖　出处：《华南农业大学》2016年硕士论文　论文类型：学位论文

【摘要】：本试验选用3T3-L1前脂肪细胞系为研究对象,探讨miR-92a-3p对3T3-L1前脂肪细胞增殖与分化的影响,旨在探讨肥胖症的发生机制,并用化学修饰模拟物miR-92a-3p mimic和miR-92a-3p inhibitor进行转染,运用CCK-8检测细胞增殖情况,细胞流式仪检测细胞周期情况,油红O染色和甘油三酯检测,实时荧光定量和蛋白免疫印迹法检测相关基因表达等方法探讨miR-92a-3p对3T3-L1前脂肪细胞细胞增殖、细胞周期、成脂分化及相关基因和蛋白表达的变化,得到以下结果:(1)过表达miR-92a-3p抑制3T3-L1前脂肪细胞增殖,而抑制miR-92a-3p表达则促进3T3-L1前脂肪细胞增殖。(2)过表达miR-92a-3p能通过下调细胞周期标志基因(Cyclin D1、Cyclin D3和CDK4)的mRNA和蛋白水平的表达,导致3T3-L1前脂肪细胞发生周期G1期阻滞进而抑制细胞增殖,而抑制miR-92a-3p表达则有相反的作用。(3)过表达miR-92a-3p在3T3-L1前脂肪细胞分化的整个过程显著上调,并呈现时间依赖关系。(4)过表达miR-92a-3p,促进脂肪细胞分化标志基因(PPARγ、C/EBPα、FABP4)的表达和甘油三酯的聚积,进而促进3T3-L1前体脂肪细胞成脂分化,而抑制miR-92a-3p表达有相反的效果。
[Abstract]:In this study, 3T3-L1 preadipose cell line was selected as the study object to investigate the effect of miR-92a-3p on the proliferation and differentiation of 3T3-L1 preadipocytes, to explore the mechanism of obesity, and to transfect 3T3-L1 preadipocytes with chemically modified mimics miR-92a-3p mimic and miR-92a-3p inhibitor. Cell proliferation was detected by CCK-8, cell cycle was detected by flow cytometry, oil red O staining and triglyceride were detected. Real-time fluorescence quantitative analysis and Western blotting were used to investigate the effects of miR-92a-3p on the proliferation, cell cycle, adipogenic differentiation and the expression of related genes and proteins in 3T3-L1 preadipocytes. The results showed that the overexpression of miR-92a-3p inhibited the proliferation of 3T3-L1 preadipocytes, while the inhibition of miR-92a-3p expression promoted the proliferation of 3T3-L1 preadipocytes. The overexpression of miR-92a-3p could inhibit the expression of mRNA and protein by down-regulating the cell cycle marker genes: Cyclin D1Cyclin D3 and CDK4). The G 1 phase arrest of 3T3-L1 preadipocytes resulted in the inhibition of cell proliferation, while the inhibition of miR-92a-3p expression had the opposite effect. The overexpression of miR-92a-3p was significantly up-regulated in the whole differentiation process of 3T3-L1 preadipocytes. The expression of miR-92a-3p was overexpressed, which promoted the expression of PPAR- 纬 -C / EBP 伪 -FABP4) and the accumulation of triglyceride, thus promoted the adipocyte adipocyte differentiation of 3T3-L1, but inhibited the expression of miR-92a-3p.
【学位授予单位】：华南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R589.2

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