Institute for Virology, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, China.
Cancer Res. 2010 Dec 1;70(23):9798-807. doi: 10.1158/0008-5472.CAN-10-1001. Epub 2010 Nov 23.
Diagnosis of hepatitis B virus (HBV)-positive hepatocellular carcinoma (HCC), particularly HCC independent of cirrhosis etiology, presents a great challenge because of a lack of biomarkers. Here we test the hypothesis that expression profiles of microRNAs (miRNAs) in serum can serve as biomarkers for diagnosis of HBV infection and HBV-positive HCC. We recruited 513 subjects (210 controls and 135 HBV-, 48 hepatitis C virus (HCV)-, and 120 HCC-affected individuals) and employed a strategy of initial screening by Solexa sequencing followed by validation with TaqMan probe-based quantitative reverse transcription-PCR assay. First, because of a close link between chronic hepatitis B and HCC, we compared miRNA expression profiles in HBV serum with that in control serum and successfully obtained 13 miRNAs that were differentially expressed in HBV serum. This 13-miRNA-based biomarker accurately discriminated not only HBV cases from controls and HCV cases, but also HBV-positive HCC cases from control and HBV cases. Second, we directly compared miRNA expressions in HCC serum with those in controls and identified 6 miRNAs that were significantly upregulated in HCC samples. Interestingly, 2 of these miRNAs, miR-375 and miR-92a, were also identified by our first approach as HBV specific. When we employed 3 of these miRNAs (miR-25, miR-375, and let-7f) as biomarkers, we could clearly separate HCC cases from controls, and miR-375 alone had an ROC of 0.96 (specificity: 96%; sensitivity: 100%) in HCC prediction. In conclusion, our study demonstrates for the first time that serum miRNA profiles can serve as novel and noninvasive biomarkers for HBV infection and HBV-positive HCC diagnosis.
乙型肝炎病毒（HBV）阳性肝细胞癌（HCC）的诊断，尤其是非肝硬化病因的 HCC，由于缺乏生物标志物而极具挑战性。在这里，我们检验了一个假说，即血清中 microRNA（miRNA）的表达谱可以作为诊断 HBV 感染和 HBV 阳性 HCC 的生物标志物。我们招募了 513 名受试者（210 名对照者和 135 名 HBV 感染者、48 名丙型肝炎病毒（HCV）感染者和 120 名 HCC 患者），并采用了索莱克斯测序初筛，然后采用 TaqMan 探针定量逆转录-PCR 验证的策略。首先，由于慢性乙型肝炎与 HCC 密切相关，我们比较了 HBV 血清和对照者血清中的 miRNA 表达谱，成功获得了 13 种在 HBV 血清中差异表达的 miRNA。这种基于 13 种 miRNA 的生物标志物不仅能准确区分 HBV 病例与对照者和 HCV 病例，还能区分 HBV 阳性 HCC 病例与对照者和 HBV 病例。其次，我们直接比较了 HCC 血清和对照者血清中的 miRNA 表达，发现 6 种 miRNA 在 HCC 样本中显著上调。有趣的是，这 6 种 miRNA 中有 2 种（miR-375 和 miR-92a）在我们的第一种方法中也被鉴定为 HBV 特异性。当我们将这 3 种 miRNA（miR-25、miR-375 和 let-7f）作为生物标志物时，可以清楚地将 HCC 病例与对照者区分开来，miR-375 单独用于 HCC 预测时的 ROC 为 0.96（特异性：96%；灵敏度：100%）。总之，本研究首次证明，血清 miRNA 谱可以作为 HBV 感染和 HBV 阳性 HCC 诊断的新型非侵入性生物标志物。
