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RNA 干扰介导的小鼠钾通道亚家族 K 成员 10 基因敲低抑制激素诱导的 3T3-L1 前脂肪细胞分化。

RNA interference-mediated knockdown of the mouse gene encoding potassium channel subfamily K member 10 inhibits hormone-induced differentiation of 3T3-L1 preadipocytes.

机构信息

National Livestock Breeding Center, Nishigo, Fukushima 961-8511, Japan.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2010 Sep;157(1):46-53. doi: 10.1016/j.cbpb.2010.04.015. Epub 2010 May 10.

Abstract

Intramuscular fat (IMF) is an economically important trait of domestic meat animals; thus, it is important to identify the factors that influence the IMF content. In this study, we identified the gene associated with adipogenesis from all the positional candidate genes located in the quantitative trait loci (QTL) for IMF content on porcine chromosome 7. We analyzed the expression of the abovementioned genes during differentiation of mouse 3T3-L1 preadipocytes by using real-time polymerase chain reaction (PCR). Total cellular RNA was extracted before and 6, 12, 36, and 48 h and 4, 6, and 8d after treatment with standard hormonal inducers of differentiation-insulin, dexamethasone, and 3-isobutyl-1-methylxanthine (IBMX). Six hours after induction, potassium channel subfamily K member 10 (KCNK10) gene expression in the preadipocytes was found to be 100-fold greater than that at the baseline; this expression declined until day 4 after the induction. Moreover, knockdown of the KCNK10 gene by transfection with short-hairpin RNA (shRNA) significantly decreased triacylglycerol accumulation on day 8 after the induction. An RNA interference study revealed that KCNK10 knockdown inhibited the differentiation of 3T3-L1 cells. These results indicate that KCNK10 plays an important role in the early stages of preadipocyte differentiation.

摘要

肌内脂肪(IMF)是家畜肉的一个重要经济性状,因此,确定影响 IMF 含量的因素非常重要。本研究从猪 7 号染色体上 IMF 含量的数量性状位点(QTL)中所有定位候选基因中鉴定出了与脂肪生成相关的基因。我们通过实时聚合酶链反应(PCR)分析了上述基因在小鼠 3T3-L1 前体脂肪细胞分化过程中的表达。在用标准激素诱导分化(胰岛素、地塞米松和 3-异丁基-1-甲基黄嘌呤(IBMX))之前以及处理后 6、12、36 和 48 小时和 4、6 和 8 天提取总细胞 RNA。诱导后 6 小时,前体脂肪细胞中的钾通道亚家族 K 成员 10(KCNK10)基因表达量比基线高 100 倍;这种表达水平一直下降到诱导后第 4 天。此外,通过短发夹 RNA(shRNA)转染敲低 KCNK10 基因显著降低了诱导后第 8 天的三酰甘油积累。RNA 干扰研究表明,KCNK10 敲低抑制了 3T3-L1 细胞的分化。这些结果表明,KCNK10 在 3T3-L1 前体脂肪细胞分化的早期阶段发挥重要作用。

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