Liver Cancer Institute and Zhongshan Hospital, Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
PLoS One. 2012;7(12):e52393. doi: 10.1371/journal.pone.0052393. Epub 2012 Dec 20.
It is still difficult to predict the probability of tumor recurrence after resection of hepatocellular carcinoma (HCC). In this study, we set out to identify specific microRNA (miRNA) in microdissected hepatitis B virus (HBV)-related HCC tissue from formalin-fixed paraffin-embedded (FFPE) samples which might be used in predicting early recurrence after HCC resection. Taqman low density arrays were used to detect the 667 miRNA profiles in both the microdissected tumorous and adjacent non-tumorous liver tissues from 20 HCC patients (discovery set) including 10 patients with early tumor recurrence and 10 without early tumor recurrence and to identify the differentially expressed miRNAs related to HCC recurrence. Then quantitative real-time PCR (qRT-PCR) was used to verify the findings in 106 patients (training set), and to develop a predictive assay. The identified miRNAs were further validated in an independent cohort of 112 patients (validation set). Thirty seven miRNAs were identified from 20 HCC patients and validated in 106 HCC patients using qRT-PCR. A significant association was found between miR-29a-5p level in HCC tissues and early tumor recurrence (P = 0.0002). This association was further confirmed in the independent validation set of 112 patients (P = 0.0154). MiR-29a-5p level was significantly associated with both time to tumor recurrence (TTR) (P = 0.0015) and overall survival (OS) (P = 0.0079) in validation set. In the multivariate analyses, miR-29a-5p was identified as an independent factor for TTR, particularly for those patients with early stage of HCC. The sensitivity and specificity of miR-29a-5p for the prediction of early HCC recurrence of BCLC 0/A stage HCC were 74.2% and 68.2%, respectively. These suggest that miR-29a-5p might be a useful marker for the prediction of early tumor recurrence after HCC resection, especially in BCLC 0/A stage HCCs.
肝癌（HCC）切除术后肿瘤复发的概率仍然难以预测。在这项研究中，我们着手从福尔马林固定石蜡包埋（FFPE）样本中微切割的乙型肝炎病毒（HBV）相关 HCC 组织中鉴定出特定的 microRNA（miRNA），这些 miRNA 可能用于预测 HCC 切除术后的早期复发。Taqman 低密度阵列用于检测来自 20 名 HCC 患者（发现集）的微切割肿瘤和相邻非肿瘤组织中的 667 种 miRNA 图谱，包括 10 名早期肿瘤复发患者和 10 名无早期肿瘤复发患者，以鉴定与 HCC 复发相关的差异表达 miRNA。然后，使用定量实时 PCR（qRT-PCR）在 106 名患者（训练集）中验证发现结果，并开发预测检测。在 112 名患者的独立队列中进一步验证了鉴定出的 miRNA（验证集）。使用 qRT-PCR 从 20 名 HCC 患者中鉴定出 37 种 miRNA，并在 106 名 HCC 患者中进行了验证。在 HCC 组织中 miR-29a-5p 水平与早期肿瘤复发之间存在显著相关性（P = 0.0002）。在 112 名独立验证患者中进一步证实了这种相关性（P = 0.0154）。miR-29a-5p 水平与验证集中的肿瘤复发时间（TTR）（P = 0.0015）和总生存（OS）（P = 0.0079）显著相关。在多变量分析中，miR-29a-5p 被确定为 TTR 的独立因素，尤其是对于 HCC 早期患者。miR-29a-5p 预测 BCLC 0/A 期 HCC 早期 HCC 复发的敏感性和特异性分别为 74.2％和 68.2％。这些表明 miR-29a-5p 可能是预测 HCC 切除术后早期肿瘤复发的有用标志物，尤其是在 BCLC 0/A 期 HCC 中。
