RNA interference-mediated knockdown of the mouse gene encoding potassium channel subfamily K member 10 inhibits hormone-induced differentiation of 3T3-L1 preadipocytes
RNA interference-mediated knockdown of the mouse gene encoding potassium channel subfamily K member 10 inhibits hormone-induced differentiation of 3T3-L1 preadipocytes
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DOI：
10.1016/j.cbpb.2010.04.015
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发表时间：
2010-09-01
影响因子：
2.2
通讯作者：
Kobayashi, Eiji
中科院分区：
文献类型：
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作者：
Sato, Shuji;Nishizuka, Makoto;Kobayashi, Eiji
肌内脂肪（IMF）是家畜的一个重要经济性状，因此，确定影响IMF含量的因素是很重要的。本研究从猪7号染色体上IMF含量的数量性状基因座（QTL）中筛选出与脂肪形成相关的基因。我们用实时荧光定量PCR技术分析了上述基因在小鼠3 T3-L1前脂肪细胞分化过程中的表达。总细胞RNA提取前和6，12，36，48小时和4，6，8天后，用标准激素诱导剂的分化胰岛素，地塞米松，3-异丁基-1-甲基黄嘌呤（IBMX）。诱导后6小时，发现前脂肪细胞中钾通道亚家族K成员10（KCNK 10）基因表达比基线时高100倍：该表达下降至诱导后第4天。此外，敲低KCNK 10基因转染短发夹RNA（shRNA）显着降低三酰甘油的积累后第8天的诱导。RNA干扰研究显示，KCNK 10基因敲低抑制3 T3-L1细胞的分化。这些结果表明，KCNK 10在前脂肪细胞分化的早期阶段起着重要作用。(C)2010年爱思唯尔公司All rights reserved.
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 8 d 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. (C) 2010 Elsevier Inc. All rights reserved.