Department of Internal Medicine, University of Rome "Tor Vergata", Via Montpellier 1, 00133 Rome, Italy.
J Gastroenterol. 2011 Dec;46(12):1391-402. doi: 10.1007/s00535-011-0456-0. Epub 2011 Aug 24.
MicroRNAs (miRNA) are tiny, noncoding, small, endogenous RNAs that play major roles in neoplastic transformation and could therefore offer a better quantitative and noninvasive method for the diagnosis and prognosis of colorectal cancer (CRC) using feces. In the present study, we screened feces for 648 miRNAs and analyzed the role of miR-144* as a potential CRC diagnostic marker.
Fecal miRNA expression was profiled with RT-pre-amplification-qPCR, and the stability was determined using both endogenous and exogenous miRNA by RT-qPCR. ROC analysis was performed to enhance the diagnosing power of the CRC patients' fecal specimens.
We detected 39% of all the miRNAs screened in feces. Endogenous miRNAs are more stable over time and temperature, while exogenous miRNAs degraded rapidly. miR-144* was overexpressed in feces, suggesting that it could be a potent candidate diagnostic marker for CRC detection, with a sensitivity of 74% and a specificity of 87% (n = 75, p < 0.0001). Moreover, RT-qPCR analysis showed that miR-144* was also overexpressed in paired CRC tissues, thus suggesting its possible utilization as a diagnostic marker.
We demonstrated that miRNAs are stable in the fecal microenvironment, and that, among them, miR-144* represents a novel fecal-based diagnostic marker for CRC screening. Nevertheless, our data need to be validated in a large cohort of subjects.
MicroRNAs(miRNA)是微小的、非编码的、小型的内源性 RNA,在肿瘤转化中起主要作用,因此可以为使用粪便进行结直肠癌(CRC)的诊断和预后提供更好的定量和非侵入性方法。在本研究中,我们筛选了粪便中的 648 个 miRNA,并分析了 miR-144*作为潜在 CRC 诊断标志物的作用。
使用 RT-预扩增-qPCR 对粪便中的 miRNA 表达进行了分析,并通过 RT-qPCR 确定了内源性和外源性 miRNA 的稳定性。ROC 分析用于增强 CRC 患者粪便标本的诊断能力。
我们检测到粪便中所有筛选出的 miRNA 的 39%。内源性 miRNA 随着时间和温度的变化更加稳定,而外源性 miRNA 则迅速降解。miR-144在粪便中表达上调,表明其可能是 CRC 检测的有效候选诊断标志物,其敏感性为 74%,特异性为 87%(n=75,p<0.0001)。此外,RT-qPCR 分析显示 miR-144在配对的 CRC 组织中也表达上调,因此提示其可能作为诊断标志物的应用。
我们证明了 miRNA 在粪便微环境中稳定存在,并且其中 miR-144*代表了一种新型的基于粪便的 CRC 筛查诊断标志物。然而,我们的数据需要在更大的研究对象队列中进行验证。