Révillion F, Pawlowski V, Hornez L, Peyrat J P
Laboratoire d'Oncologie Moléculaire Humaine, Centre Oscar Lambret, Lille, France.
Eur J Cancer. 2000 May;36(8):1038-42. doi: 10.1016/s0959-8049(00)00051-4.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been widely used as a control RNA in Northern blotting and in reverse transcriptase-polymerase chain reaction (RT-PCR) analyses. We investigated the expression of GAPDH in a large series of primary breast cancers and in MCF7 human mammary epithelial breast cancer cells treated with oestradiol. The expression of GAPDH was quantified by a real-time one-step RT-PCR assay, based upon the 5' nuclease activity of Taq polymerase using an Abi Prism 7700 Sequence Detector System (Perkin Elmer, France). Using the Spearman test, GAPDH expression was found to correlate inversely with the age of the patients at diagnosis (P = 0.003; r = -0.147), oestradiol receptors (ER) (P<0.0001; r = -0.327) and progesterone receptors (PgR) (P < 0.0001; r = -0.206). A positive correlation was observed between GAPDH expression and the histo-prognostic grading (HPG) (P < 0.0001; r = 0.344). Moreover, the overall survival (OS) and the relapse-free survival (RFS) were significantly reduced in patients whose tumours showed an enhanced level of GAPDH expression (OS, P = 0.046; RFS, P = 0.021). Multivariate analyses demonstrated that GAPDH was not an independent prognostic factor. Finally, in MCF7 cells treated with oestradiol. a statistically significant dose-dependent increase in GAPDH expression was observed. These results show that GAPDH expression is associated with breast cancer cell proliferation and with the aggressiveness of tumours. The present study demonstrates that, in cancer, the use of GAPDH gene expression should not be used as a control RNA.
甘油醛-3-磷酸脱氢酶（GAPDH）已被广泛用作Northern印迹分析以及逆转录聚合酶链反应（RT-PCR）分析中的对照RNA。我们研究了GAPDH在一系列原发性乳腺癌以及经雌二醇处理的MCF7人乳腺上皮癌细胞中的表达情况。采用实时一步法RT-PCR测定法，基于Taq聚合酶的5'核酸酶活性，使用Abi Prism 7700序列检测系统（法国珀金埃尔默公司）对GAPDH的表达进行定量分析。通过Spearman检验发现，GAPDH表达与患者诊断时的年龄呈负相关（P = 0.003；r = -0.147），与雌激素受体（ER）呈负相关（P<0.0001；r = -0.327），与孕激素受体（PgR）呈负相关（P < 0.0001；r = -0.206）。观察到GAPDH表达与组织预后分级（HPG）呈正相关（P < 0.0001；r = 0.344）。此外，肿瘤中GAPDH表达水平升高的患者其总生存期（OS）和无复发生存期（RFS）显著缩短（OS，P = 0.046；RFS，P = 0.021）。多变量分析表明，GAPDH不是一个独立的预后因素。最后，在经雌二醇处理的MCF7细胞中，观察到GAPDH表达呈统计学上显著的剂量依赖性增加。这些结果表明，GAPDH表达与乳腺癌细胞增殖以及肿瘤侵袭性相关。本研究表明，在癌症研究中，不应将GAPDH基因表达用作对照RNA。
