Department of Radiotherapy, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, China.
Mol Biol Rep. 2013 Aug;40(8):5027-34. doi: 10.1007/s11033-013-2603-6. Epub 2013 Jun 21.
MicroRNAs are endogenous small RNAs with a high degree of conservation, participating in a variety of vital activities. In present study, to explore the effect of microRNAs on 3T3-L1 adipocyte differentiation and adiponectin expression, the adipo-related microRNAs were screened and identified by micorRNA microarray. The highly expression plasmid of microRNA-21 with obvious expression up-regulation (miR-21) and its anti-sense (miR-21 inhibitor) were constructed and transfected into 3T3-L1 preadipocytes. The effect of miR-21 on 3T3-L1 adipocyte differentiation was observed, and the protein and mRNA expression level of adiponectin and AP-1 were analyzed. Results showed that, the expression profiles of microRNAs significantly changed during 3T3-L1 adipocyte differentiation. The expression of miR-21 was obviously up-regulated. miR-21 could significantly promote adipocyte differentiation, increase adiponectin mRNA and protein expression, while decrease AP-1 protein level. Meanwhil, miR-21 inhibitor blocked the effects of miR-21 mentioned above. The overexpression of AP-1 could absolutely reverse the stimulatory effect of miR-21 on adiponectin. miR-21 plays an important role in regulating adipocyte differentiation and adiponectin expression by inhibiting AP-1 expression.
微小 RNA 是具有高度保守性的内源性小 RNA,参与多种重要的生命活动。本研究通过 miRNA 微阵列筛选和鉴定与脂肪相关的 microRNAs,探讨 microRNAs 对 3T3-L1 脂肪细胞分化和脂联素表达的影响。构建了微小 RNA-21 高表达质粒(miR-21)及其反义(miR-21 抑制剂),并转染至 3T3-L1 前脂肪细胞。观察 miR-21 对 3T3-L1 脂肪细胞分化的影响,分析脂联素和 AP-1 的蛋白和 mRNA 表达水平。结果表明,微小 RNA 在 3T3-L1 脂肪细胞分化过程中的表达谱发生明显变化。miR-21 的表达明显上调。miR-21 可显著促进脂肪细胞分化,增加脂联素 mRNA 和蛋白表达,同时降低 AP-1 蛋白水平。同时,miR-21 抑制剂阻断了上述 miR-21 的作用。AP-1 的过表达可完全逆转 miR-21 对脂联素的刺激作用。miR-21 通过抑制 AP-1 的表达在调节脂肪细胞分化和脂联素表达中起重要作用。