Effects of dimethyl sulfoxide and dexamethasone on mRNA expression of housekeeping genes in cultures of C2C12 myotubes
Effects of dimethyl sulfoxide and dexamethasone on mRNA expression of housekeeping genes in cultures of C2C12 myotubes
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DOI：
10.1016/j.bbrc.2008.01.006
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发表时间：
2008-03-14
影响因子：
3.1
通讯作者：
Ikeda, Muneharu
中科院分区：
文献类型：
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作者：
Nishimura, Masuhiro;Nikawa, Takeshi;Ikeda, Muneharu
我们采用实时荧光定量RT-PCR技术研究了二甲基亚砜（DMSO）和地塞米松（Dex）对管家基因β-肌动蛋白（ACTB）、甘油醛-3-磷酸脱氢酶（GAPDH）、β-葡萄糖醛酸酶（GUSB）、次黄嘌呤磷酸核糖转移酶1（HPRT 1）、磷酸甘油酸激酶1（PGK 1）、肽基脯氨酰异构酶A（PPIA）、和转铁蛋白受体（TFRC）。在从成肌细胞分化为肌管的C2 C12细胞中，ACTB mRNA水平与HPRT 1 mRNA水平的比率从培养的0至120小时降低，而TFRC mRNA水平与HPRT 1 mRNA水平的比率从培养的0至120小时增加。GAPDH、GUSB、PGK 1和PPIA mRNA水平与HPRT 1 mRNA水平的比率在培养的0至120 h保持恒定。所有管家基因的mRNA水平不受暴露于0.1%或更低浓度的DMSO的影响。GAPDH mRNA水平被Dex增加，而ACTB和PGK 1 mRNA水平被Dex显著降低。GUSB、PPIA和TFRC mRNA水平不受Dex暴露的影响。因此，GUSB、HPRT 1和PPIA是用于评价C2 C12细胞培养物中mRNA表达水平的合适内部对照。(C)2008年爱思唯尔公司All rights reserved.
We used quantitative real-time RT-PCR to investigate the effects of dimethyl sulfoxide (DMSO) and dexamethasone (Dex) on the mRNA expression levels of the housekeeping genes beta-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-glucuronidase (GUSB), hypoxanthine phosphoribosyltransferase 1 (HPRT1), phosphoglycerate kinase 1 (PGK1), peptidylprolyl isomerase A (PPIA), and transferrin receptor (TFRC) in cultures of C2C12 myotubes. The ratios of ACTB mRNA levels to the HPRT1 mRNA level in C2C12 cells that were differentiating from myoblast cells to myotubes decreased from 0 to 120 h of culture, whereas the ratios of TFRC rnRNA levels to the HPRT1 mRNA level increased from 0 to 120 h of culture. The ratios of GAPDH, GUSB, PGK1, and PPIA mRNA levels to the HPRT1 mRNA level remained constant from 0 to 120 h of culture. All housekeeping gene mRNA levels were unaffected by exposure to DMSO concentrations of 0.1% or less. The GAPDH mRNA level was increased by Dex, while the ACTB and PGK1 mRNA levels were significantly decreased by Dex. The GUSB, PPIA, and TFRC mRNA levels were unaffected by exposure to Dex. GUSB, HPRT1, and PPIA are thus suitable internal controls for evaluating mRNA expression levels in cultures of C2C12 cells. (C) 2008 Elsevier Inc. All rights reserved.