Department of Neurosurgery, Sylvester Comprehensive Cancer Center, University of Miami Brain Tumor Initiative, University of Miami Miller School of Medicine, Miami, Florida, 33136, USA.
Department of Psychiatry and Behavioral Sciences, Center for Therapeutic Innovation, Miami Project to Cure Paralysis, Sylvester Comprehensive Cancer Center, University of Miami Brain Tumor Initiative, University of Miami Miller School of Medicine, Miami, Florida, 33136, USA.
Mol Cancer. 2018 Mar 20;17(1):74. doi: 10.1186/s12943-018-0822-0.
Glioblastoma multiforme (GBM) is the most common and aggressive malignant adult primary brain tumor. Despite surgical resection followed by radiotherapy and chemotherapy, the median survival rate is approximately 14 months. Although experimental therapies are in clinical trials for GBM, there is an urgent need for a peripheral GBM biomarker for measuring treatment response. As we have previously demonstrated that the long noncoding RNA HOX Transcript Antisense Intergenic RNA, or HOTAIR, is dysregulated in GBM and required for GBM cell proliferation, we hypothesized that HOTAIR expression may be utilized as a peripheral biomarker for GBM. HOTAIR expression was measured in serum from 43 GBM and 40 controls using quantitative real-time PCR (qRT-PCR). The PCR products were subsequently subcloned into pCR™4-TOPO®TA vectors for DNA sequencing. A ROC curve was also generated to examine HOTAIR's prognostic value. The amount of HOTAIR in serum exosomes and exosome-depleted supernatant was calculated by qRT-PCR. The relative HOTAIR expression was also investigated in 15 pairs of GBM serum and tumors. We detected HOTAIR in serum from GBM patients. HOTAIR levels in serum samples from GBM patients was significantly higher than in the corresponding controls (P < 0.0001). The area under the ROC curve distinguishing GBM patients from controls was 0.913 (95% CI: 0.845-0.982, P < 0.0001), with 86.1% sensitivity and 87.5% specificity at the cut-off value of 10.8. HOTAIR expression was significantly correlated with high grade brain tumors. In addition, Pearson correlation analysis indicated a medium correlation of serum HOTAIR levels and the corresponding tumor HOTAIR levels (r = 0.734, P < 0.01). We confirmed via sequencing that the amplified HOTAIR from serum contained the HOTAIR sequence and maps to the known HOTAIR locus at 12q13. The serum-derived exosomes contain HOTAIR and the purified exosomes were validated by western blot and nanoparticle tracking analysis. Importantly, our results demonstrate that serum HOTAIR can be used as a novel prognostic and diagnostic biomarker for GBM.
多形性胶质母细胞瘤（GBM）是最常见和侵袭性最强的成人原发性脑肿瘤。尽管进行了手术切除，然后进行放疗和化疗，但中位生存率约为 14 个月。尽管实验疗法正在进行临床试验，但迫切需要一种外周 GBM 生物标志物来测量治疗反应。正如我们之前所证明的那样，长链非编码 RNA HOX 转录反义基因间 RNA（或 HOTAIR）在 GBM 中失调，并且是 GBM 细胞增殖所必需的，因此我们假设 HOTAIR 表达可用作 GBM 的外周生物标志物。使用实时定量 PCR（qRT-PCR）测量了来自 43 名 GBM 和 40 名对照者的血清中的 HOTAIR 表达。随后将 PCR 产物亚克隆到 pCR™4-TOPO®TA 载体中进行 DNA 测序。还生成了 ROC 曲线以检查 HOTAIR 的预后价值。通过 qRT-PCR 计算血清外泌体和外泌体耗尽上清液中的 HOTAIR 量。还研究了 15 对 GBM 血清和肿瘤中 HOTAIR 的相对表达。我们在 GBM 患者的血清中检测到了 HOTAIR。GBM 患者血清样本中的 HOTAIR 水平明显高于相应的对照者（P <0.0001）。区分 GBM 患者和对照者的 ROC 曲线下面积为 0.913（95%CI：0.845-0.982，P <0.0001），截断值为 10.8 时，灵敏度为 86.1%，特异性为 87.5%。HOTAIR 表达与高级别脑肿瘤显着相关。此外，Pearson 相关分析表明血清 HOTAIR 水平与相应肿瘤 HOTAIR 水平呈中度相关（r = 0.734，P <0.01）。我们通过测序证实，从血清中扩增的 HOTAIR 包含 HOTAIR 序列，并映射到已知的 12q13 处的 HOTAIR 基因座。血清衍生的外泌体包含 HOTAIR，并且通过 Western blot 和纳米颗粒跟踪分析验证了纯化的外泌体。重要的是，我们的结果表明血清 HOTAIR 可用作 GBM 的新型预后和诊断生物标志物。
