Sakamoto Yuko, Nishioka Junji, Noma Katsura, Yasuda Kazunari, Nakatani Kaname, Kise Hideaki, Sugimura Yoshiki, Nobori Tsutomu
Personalized Medicine/Clinical Laboratories, Mie University Hospital, Tsu 514-8507, Japan.
Rinsho Byori. 2011 May;59(5):446-51.
Survivin is one of the apoptosis inhibitor proteins and is rarely expressed in adult normal tissues. However, survivin expression has been detected in various tumors. In this study, we evaluated the usefulness of urinary survivin/glyceraldehyde-3-phosphate dehydrogenase (GAPDH) ratio as a marker for bladder tumor.
Urine samples were obtained from 72 patients with bladder tumor, 36 with urinary tract inflammation as controls. Survivin and GAPDH mRNA expression was measured by quantitative real-time PCR assay in urine cells. The GAPDH housekeeping gene was used for normalization of survivin expression. We also analyzed survivin protein levels using urine samples and recombinant protein by western blotting.
High expression of survivin was confirmed on the protein level using urine samples of bladder tumor by western blotting. Survivin/GAPDH mRNA ratios of bladder tumor quantified by real-time PCR was significantly higher than those of controls (p=0.001). In pathological stage of bladder tumor, survivin/ GAPDH mRNA ratio of pTis was significantly high compared with pTa and pT1 (p < 0.001, p=0.001, respectively). Grade3 tumors expressed high level of survivin/GAPDH mRNA ratio compared with Grade 1 and Grade 2 tumors (p=0.03). The sensitivity, the specificity and AUC(area under the curve) of survivin/ GAPDH mRNA ratio was 83.3%, 86.1% and 0.898, respectively.
Measuring survivin/GAPDH mRNA ratio in urine is non-invasive and high sensitive examination. Therefore, survivin/GAPDH mRNA ratio is useful marker for the detection of bladder tumor, especially to detect carcinoma in situ.
生存素是凋亡抑制蛋白之一,在成人正常组织中很少表达。然而,在各种肿瘤中均检测到生存素的表达。在本研究中,我们评估了尿生存素/甘油醛-3-磷酸脱氢酶(GAPDH)比值作为膀胱肿瘤标志物的实用性。
收集72例膀胱肿瘤患者的尿液样本,36例尿路炎症患者的尿液样本作为对照。采用定量实时PCR法检测尿细胞中生存素和GAPDH mRNA的表达。以GAPDH管家基因对生存素表达进行标准化。我们还通过蛋白质印迹法,利用尿液样本和重组蛋白分析了生存素蛋白水平。
通过蛋白质印迹法,利用膀胱肿瘤患者的尿液样本在蛋白水平证实了生存素的高表达。实时PCR定量检测膀胱肿瘤的生存素/GAPDH mRNA比值显著高于对照组(p = 0.001)。在膀胱肿瘤的病理分期中,pTis的生存素/GAPDH mRNA比值显著高于pTa和pT1(分别为p < 0.001,p = 0.001)。3级肿瘤的生存素/GAPDH mRNA比值高于1级和2级肿瘤(p = 0.03)。生存素/GAPDH mRNA比值的敏感性、特异性和曲线下面积(AUC)分别为83.3%、86.1%和0.898。
检测尿液中生存素/GAPDH mRNA比值是一种无创且高灵敏度的检查方法。因此,生存素/GAPDH mRNA比值是检测膀胱肿瘤,尤其是原位癌的有用标志物。