MiR-26a靶向调节PTEN对3T3-L1前脂肪细胞分化的促进效应

发布时间：2019-06-03 18:21

【摘要】：miRNA作为内源性非编码小RNA,主要在转录后水平调控基因表达。近年来越来越多的证据表明,miRNA通过调控脂肪细胞分化相关的转录因子和信号通路来影响脂肪细胞的分化。为探究miR-26a对小鼠(Mus musculus)3T3-L1前脂肪细胞分化的影响及具体的调节作用,本研究分别用miR-26a的agomir和antagomir对miR-26a进行过表达和敲除。结果显示,miR-26a的表达量在3T3-L1细胞分化过程中逐渐上升。过表达miR-26a极显著增加了脂肪合成相关基因——过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor gamma,PPARγ)、脂肪酸合酶(fatty acid synthase,FAS)和脂蛋白脂酶(lipoprotein lipase,LPL)的mRNA水平(P0.01),PPARγ和FAS的蛋白水平也显著升高(P0.05)。同时,细胞内的脂质聚集也明显增加。相反,抑制miR-26a的表达量降低了3T3-L1细胞的分化。通过靶基因预测和双荧光素酶报告系统分析,证明miR-26a能直接结合到同源性磷酸酶-张力蛋白(phosphatase and tensin homolog,PTEN)mRNA的3'UTR,并且导致其miRNA和蛋白水平降低(P0.05)。本研究的结果表明,miR-26a可能通过直接抑制PTEN来正向的调节3T3-L1细胞的脂肪分化,该结果为miRNA成为治疗肥胖相关疾病的药物靶分子提供了一定的理论依据。
[Abstract]:MiRNA, as an endogenous non-coding small RNA, mainly regulates gene expression at the post-transcriptional level. In recent years, more and more evidence suggests that miRNA affects adipocytes differentiation by regulating transcription factors and signal pathways related to adipocytes differentiation. In order to investigate the effect of miR-26a on the differentiation of mouse (Mus musculus) 3T3-L1 preadipocytes and its regulatory effect, miR-26a was overexpressed and knockout by agomir and antagomir of miR-26a, respectively. The results showed that the expression of miR-26a increased gradually during the differentiation of 3T3-L1 cells. Overexpression of miR-26a significantly increased peroxisome proliferator activated receptor gamma (peroxisome proliferator-activated receptor gamma,PPAR 纬), fatty acid synthase (fatty acid synthase,FAS and lipoprotein lipase (lipoprotein lipase, which are related to fat synthesis. The mRNA level of LPL (P 0.01), the protein level of PPR 纬 and FAS were also significantly increased (P 0.05). At the same time, the accumulation of lipid in cells was also significantly increased. On the contrary, inhibition of miR-26a expression decreased the differentiation of 3T3-L1 cells. Through target gene prediction and double luciferase reporting system analysis, it was proved that miR-26a could directly bind to 3 UTRs of homology phosphatase-tensin (phosphatase and tensin homolog,PTEN mRNA, and lead to the decrease of miRNA and protein levels (P 0.05). The results of this study suggest that miR-26a may positively regulate the adipose differentiation of 3T3-L1 cells by directly inhibiting PTEN, which provides a theoretical basis for miRNA to become a drug target molecule for the treatment of obesity-related diseases.
【作者单位】：四川农业大学动物遗传育种研究所;四川农业大学动物科技学院;
【基金】：国家高技术研究发展计划(863计划)(No.2013AA102502) 国家自然科学基金(No.31472081和No.31522055)
【分类号】：R589.2

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