Iempridee Tawin, Wiwithaphon Suphachai, Piboonprai Kitiya, Pratedrat Pornpitra, Khumkhrong Phattharachanok, Japrung Deanpen, Temisak Sasithon, Laiwejpithaya Somsak, Chaopotong Pattama, Dharakul Tararaj
National Nanotechnology Center National Science and Technology Development Agency Pathum Thani Thailand.
National Institute of Metrology Pathum Thani Thailand.
FEBS Open Bio. 2018 Sep 28;8(11):1844-1854. doi: 10.1002/2211-5463.12523. eCollection 2018 Nov.
Circulating lncRNAs have attracted considerable attention as potential noninvasive biomarkers for diagnosing cancers. RT-qPCR is the canonical technique for detecting circulating RNA and depends largely on stable reference genes for data normalization. However, no systematic evaluation of reference genes for serum lncRNA has been reported for cervical cancer. Here, we profiled and validated lncRNA expression from serum of cervical cancer patients and controls using microarrays and RT-qPCR. We identified lncRNA RP11-204K16.1, XLOC_012542, and U6 small nuclear RNA as the most stable reference genes based on geNorm, NormFinder, BestKeeper, delta Ct, and RefFinder. These genes were suitable also for samples from different age groups or with hemolysis. Additionally, we discovered lncRNA AC017078.1 and XLOC_011152 as candidate biomarkers, whose expression was down-regulated in cervical cancer. Our findings could aid research on circulating lncRNA and the discovery of blood-based biomarkers for cervical cancer diagnosis.
循环长链非编码RNA（lncRNAs）作为诊断癌症的潜在非侵入性生物标志物已引起广泛关注。逆转录定量聚合酶链反应（RT-qPCR）是检测循环RNA的经典技术，很大程度上依赖稳定的内参基因进行数据标准化。然而，尚未见针对宫颈癌血清lncRNA内参基因的系统评估报道。在此，我们使用微阵列和RT-qPCR分析并验证了宫颈癌患者和对照血清中的lncRNA表达。基于geNorm、NormFinder、BestKeeper、delta Ct和RefFinder，我们确定lncRNA RP11-204K16.1、XLOC_012542和U6小核RNA为最稳定的内参基因。这些基因也适用于不同年龄组或有溶血的样本。此外，我们发现lncRNA AC017078.1和XLOC_011152为候选生物标志物，其表达在宫颈癌中下调。我们的研究结果有助于循环lncRNA的研究以及发现用于宫颈癌诊断的血液生物标志物。
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