miR-26a-5p对3T3-L1前脂肪细胞增殖和分化的影响

Effects of miR-26a-5p on proliferation and differentiation of 3T3-L1 preadipocyte

分别将miR-26a-5p mimic和inhibitor成功转染至3T3-L1前脂肪细胞,使miR-26a-5p过表达和功能缺失。采用CCK-8和流式细胞术检测细胞活力及细胞周期,实时qPCR和Western blot技术分别检测细胞周期G1期标志基因CyclinD1和CDK4及成脂标志基因FABP4和C/EBPα的表达,油红O染色检测甘油三酯含量变化。结果显示：过表达miR-26a-5p显著降低3T3-L1前脂肪细胞的活力并导致其G1期阻滞;与对照组相比,miR-26a-5p mimic组CyclinD1和CDK4的mRNA和蛋白水平表达显著下调而FABP4和C/EBPα的mRNA和蛋白水平表达显著上调,甘油三酯积累增加;当抑制其表达后出现与之完全相反的结果。结果表明：miR-26a-5p显著抑制3T3-L1前脂肪细胞增殖并促进其成脂分化。

To investigate the effect of miR-26a-5p on 3T3 L1 preadipocytes,in this study, miR-26a 5p mimic and inhibitor were successfully transfected into 3T3-L1 preadipocytes to induce miR-26a- 5p overexpression and loss of function. Cell viability and cell cycle were detected by CCK-8 and flow cytometry. The expression of cell cycle genes CyclinD1, CDK4 and adipogenic marker genes C/EBPa,FABP4 was detected by real-time qPCR and Western blots. Oil red O staining was used to detect the changed triglyceride content. The results showed that overexpression of miR-26a-Sp significantly reduced the activity of 3T3-L1 preadipocytes and led to G1 phase arrest; Compared with the control group,the mRNA and protein levels of CyclinD1 and CDK4 in miR-26a-5p mimic group were significantly down-regulated, while the expression levels of mRNA and protein of FABP4 and C/EBPa were significantly up-regulated, as well as the trigIyceride accumulation increased. When the expression of miR-26a-Sp was suppressed,the opposite result was found. These results indicated that miR-26a-5p significantly inhibited the proliferation of 3T3-L1 preadipocytes and promoted their adipogenic differentiation.