Gene expression as possible biomarker in bladder urothelium of patients with ulcerative interstitial cystitis

我们通过DNA微阵列分析和实时定量聚合酶链式反应研究了溃疡性间质性膀胱炎的相关基因。取9例溃疡性间质性膀胱炎患者的膀胱尿路上皮组织和9例正常对照的膀胱尿路上皮组织，其中膀胱癌7例，良性前列腺增生症2例。从膀胱样本中提取总RNA，并使用全人类基因组DNA微阵列44K(Agilent Technologies，Santa Clara，California)比较这两组患者的基因表达。微阵列数据用GeneSpring GX软件和Invenity Path分析软件进行分析。所选基因通过实时定量聚合酶链式反应确认为转录改变。通过火山图谱分析，我们确定了564个在基因中显著表达的探针，是对照组的4倍以上(p&lt；0.001)。对这些基因的进一步网络独创性路径分析显示了最重要的3个功能，包括1)细胞间的信号和相互作用，以及血液系统的发育和功能，2)炎症性疾病和3)细胞发育。实时荧光定量聚合酶链式反应证实了CXCR3结合趋化因子(CXCL9、10、11)和肿瘤坏死因子配体超家族成员14等多种基因在溃疡性间质性膀胱炎患者膀胱组织中的高表达，这些基因可能是间质性膀胱炎的潜在生物标志物。

We investigated the genes responsible for ulcerative interstitial cystitis by DNA microarray analysis and quantitative real-time polymerase chain reaction. Bladder urothelial tissues were taken from a site apart from the ulcerative lesion in 9 patients with ulcerative interstitial cystitis and from a normal-looking area in 9 controls, including 7 with bladder carcinoma and 2 with benign prostatic hyperplasia. Total RNA was extracted from bladder samples and gene expression was compared between these 2 groups using Whole Human Genome DNA microarray 44K (Agilent Technologies, Santa Clara, California). Microarray data were analyzed by GeneSpring GX software and Ingenuity Pathway Analysis. Chosen genes were confirmed for altered transcription by quantitative real-time polymerase chain reaction. We identified 564 probes that were significantly expressed in mRNA more than 4-fold vs those in controls using volcano plot analysis (p<0.001). Further network Ingenuity Pathway Analysis of these genes showed the top 3 functions, including 1) cell-to-cell signaling and interaction, and hematological system development and function, 2) inflammatory disease and 3) cellular development. Quantitative real-time polymerase chain reaction confirmed increased mRNA expression of several genes in the bladder samples of patients with ulcerative interstitial cystitis, including CXCR3 binding chemokines (CXCL9, 10 and 11) and tumor necrosis factor ligand superfamily member 14. Our study using DNA microarray analysis followed by quantitative real-time polymerase chain reaction reveals over expression of genes related to immune and inflammatory responses, including T-helper type 1 related chemokines, and cytokines such as CXCR3 binding chemokines and tumor necrosis factor ligand superfamily member 14. These genes may be potential interstitial cystitis biomarkers.