通过RNA干扰敲低TCAP可抑制培养的骨骼肌细胞中的成肌细胞分化。
TCAP knockdown by RNA interference inhibits myoblast differentiation in cultured skeletal muscle cells.
作者信息
Markert Chad D, Ning Jie, Staley Jerry T, Heinzke Laura, Childers Charles K, Ferreira J Andries, Brown Marybeth, Stoker Aaron, Okamura Carol, Childers Martin K
机构信息
Department of Neurology and Wake Forest Institute for Regenerative Medicine, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
出版信息
Neuromuscul Disord. 2008 May;18(5):413-22. doi: 10.1016/j.nmd.2008.03.010. Epub 2008 Apr 28.
Null mutation of titin-cap (TCAP) causes limb-girdle muscular dystrophy type 2G (LGMD2G). LGMD2G patients develop muscle atrophy, and lose the ability to walk by their third decade. Previous findings suggest that TCAP regulates myostatin, a key regulator of muscle growth. We tested the hypothesis that TCAP knockdown with RNA interference will lead to differential expression of genes involved in muscle proliferation and differentiation, impairing muscle cell growth. mRNA from cultured cells treated with TCAP siRNA duplex constructs was analyzed using Northern blots and real-time RT-PCR. siRNA treatment decreased TCAP mRNA expression in differentiating muscle cells. Significant (p<0.05) decreases in mRNA were observed for myogenic regulatory factors. siRNA treatment also prevented development of the normal phenotype of muscle cells. Our findings suggest that TCAP knockdown with RNA interference alters normal muscle cell differentiation.
肌联蛋白帽（TCAP）的无效突变会导致2G型肢带型肌营养不良症（LGMD2G）。LGMD2G患者会出现肌肉萎缩，并在三十多岁时丧失行走能力。先前的研究结果表明，TCAP可调节肌肉生长的关键调节因子——肌肉生长抑制素。我们检验了以下假设：通过RNA干扰敲低TCAP会导致参与肌肉增殖和分化的基因表达差异，从而损害肌肉细胞生长。使用Northern印迹法和实时逆转录-聚合酶链反应（RT-PCR）分析了用TCAP小干扰RNA（siRNA）双链体构建体处理的培养细胞的信使核糖核酸（mRNA）。siRNA处理降低了分化中的肌肉细胞中TCAP mRNA的表达。对于生肌调节因子，观察到mRNA有显著（p<0.05）下降。siRNA处理还阻止了肌肉细胞正常表型的形成。我们的研究结果表明，通过RNA干扰敲低TCAP会改变正常的肌肉细胞分化。
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