摘要
目的探讨含钾通道四聚化结构域15(KCTD15)基因在3T3-L1脂肪前体细胞分化过程中的作用。方法①采用半定量逆转录PCR检测在3T3-L1脂肪前体细胞分化过程中KCTD15mRNA表达变化。②在3T3-L1脂肪前体细胞增殖早期通过RNA干扰技术靶向敲低KCTD15基因的表达,在靶向敲低KCTD15基因后的转染KCTD15siRNA48h后通过半定量逆转录PCR验证KCTD15基因的敲低效果。用油红O染色法观察KCTD15敲低后3T3-L1细胞第0天和第10天的细胞形态学改变。③采用半定量逆转录PCR检测KCTD15基因敲低后PPARγ、C/EBPα、C/EBPβ、C/EBPδ成脂基因的变化。结果在3T3-L1脂肪前体细胞分化过程中,KCTD15mRNA表达水平逐渐降低(P<0.05);KCTD15敲低能显著抑制3T3-L1脂肪前体细胞分化;KCTD15敲低后PPARγ、C/EBPα、C/EBPβ、C/EBPδ成脂基因无明显变化。结论在分化早期阶段敲低KCTD15基因能抑制3T3-L1脂肪前体细胞分化。
Objective To study the effect of potassium channel tetramerisation domain containing 15 （KCTD15） gene on preadipocyte differentiation. Methods The expression of KCTD15 gene during 3T3-L1 preadipocyte differentiation was detected by semi-quantitative reverse transcriptase PCR. After transferring KCTD15 siRNA into the preadipocytes, the cell morphology was observed during preadipocyte differentiation by oil red O staining, and the level of triglyceride was examined by assay kit. The expression of adipogenesis genes, peroxisome proliferator-activated receptor （PPAR）γ, CCAAT/enhancer-binding protein （C/EBP）α, C/EBPβ and C/EBPδ was detected by semi-quantitative reverse transcriptase PCR. Results The expression of KCTD15 gene was decreased during 3T3-L1 cell differentiation. KCTD15 gene knockdown inhibited the differentiation and lipid accumulation of 3T3-L1 cells, and there was no significant change in the expression of PPARγ, C/EBPα, C/EBPβ and C/EBPδ. Conclusion KCTD15 gene deficiency leads to the inhibition of 3T3-L1 preadipocyte differentiation at early stage.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第11期1115-1118,共4页
Journal of Third Military Medical University
