Identification of differentially expressed long non-coding RNAs in bladder cancer

As various data suggest that lncRNAs are important in human cancer, we hypothesized that they play a role in urothelial carcinogenesis. To test this hypothesis we analysed 83 normal and malignant urothelial samples using NCode™ Human Non-coding RNA Microarrays for differentially expressed lncRNAs with potential regulatory functions in urothelial cell cancer (UCC). We identified global changes in lncRNA expression that reflect the disease phenotype. We then performed explorative analysis (siRNA gene knock-down and loss-of-function studies) of selected species and identified those with potential prognostic roles. Quantitative real-time PCR was used to validate the microarray findings. Total RNA was extracted from urothelial samples from 57 patients with UCC (23 low-grade NMI, 14 high-grade NMI and 20 invasive tumours) and from 26 disease free controls.

鉴于多项研究数据表明长链非编码RNA（long non-coding RNA, lncRNA）在人类癌症中发挥关键作用，我们推测其同样参与尿路上皮癌变过程。为验证该假说，我们采用NCode™人类非编码RNA微阵列，对83例正常与恶性尿路上皮样本开展分析，以筛选在尿路上皮细胞癌（Urothelial Cell Cancer, UCC）中具备潜在调控功能的差异表达长链非编码RNA。本研究鉴定出了可反映疾病表型的长链非编码RNA全局表达变化谱。随后我们对筛选出的候选长链非编码RNA分子开展探索性分析（包括小干扰RNA（small interfering RNA, siRNA）基因敲降与功能丧失实验），并鉴定出具备潜在预后价值的分子。我们采用实时定量聚合酶链式反应对微阵列实验结果进行了验证。本研究从57例尿路上皮细胞癌患者（含23例低级别非肌层浸润性肿瘤、14例高级别非肌层浸润性肿瘤及20例侵袭性肿瘤）的尿路上皮样本，以及26例健康对照样本中提取了总RNA。