RATIONALE Hepatocellular carcinoma (HCC) is a highly malignant disease for which the development of prospective or prognostic biomarkers is urgently required. Although metabolomics is widely used for biomarker discovery, there are some bottlenecks regarding the comprehensiveness of detected features, reproducibility of methods, and identification of metabolites. In addition, information on localization of metabolites in tumor tissue is needed for functional analysis. Here, we developed a wide-polarity global metabolomics (G-Met) method, identified HCC biomarkers in human liver samples by high-definition mass spectrometry (HDMS), and demonstrated localization in cryosections using desorption electrospray ionization MS imaging (DESI-MSI) analysis. METHODS Metabolic profiling of tumor (n = 38) and nontumor (n = 72) regions in human livers of HCC was performed by an ultrahigh-performance liquid chromatography quadrupole time-of-flight MS (UHPLC/QTOFMS) instrument equipped with a mixed-mode column. The HCC biomarker candidates were extracted by multivariate analyses and identified by matching values of the collision cross section and their fragment ions on the mass spectra obtained by HDMS. Cryosections of HCC livers, which included both tumor and nontumor regions, were analyzed by DESI-MSI. RESULTS From the multivariate analysis, m/z 904.83 and m/z 874.79 were significantly high and low, respectively, in tumor samples and were identified as triglyceride (TG) 16:0/18:1(9Z)/20:1(11Z) and TG 16:0/18:1(9Z)/18:2(9Z,12Z) using the synthetic compounds. The TGs were clearly localized in the tumor or nontumor areas of the cryosection. CONCLUSIONS Novel biomarkers for HCC were identified by a comprehensive and reproducible G-Met method with HDMS using a mixed-mode column. The combination analysis of UHPLC/QTOFMS and DESI-MSI revealed that the different molecular species of TGs were associated with tumor distribution and were useful for characterizing the progression of tumor cells and discovering prospective biomarkers.

中文翻译：

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通过高清质谱鉴定肝细胞癌的新生物标记物：超高效液相色谱四极杆飞行时间质谱和解吸电喷雾电离质谱成像。

理由肝细胞癌（HCC）是一种高度恶性的疾病，因此迫切需要开发前瞻性或预后性生物标志物。尽管代谢组学被广泛用于生物标志物的发现，但是关于检测到的特征的全面性，方法的可重复性以及代谢物的鉴定仍存在一些瓶颈。此外，功能分析还需要有关肿瘤组织中代谢物定位的信息。在这里，我们开发了一种宽极性全球代谢组学（G-Met）方法，通过高清质谱（HDMS）在人类肝脏样品中鉴定了HCC生物标志物，并使用解吸电喷雾电离MS成像（DESI-MSI）展示了冰冻切片中的定位。分析。方法采用配备了高效液相色谱四极杆飞行时间质谱仪（UHPLC / QTOFMS）的超高效液相色谱四极杆飞行时间质谱仪（UHPLC / QTOFMS）对HCC人肝中肿瘤（n = 38）和非肿瘤（n = 72）区域进行代谢谱分析模式列。通过多变量分析提取HCC生物标志物候选物，并通过HDMS获得的质谱图上的碰撞截面及其碎片离子的匹配值进行识别。通过DESI-MSI分析了包括肿瘤和非肿瘤区域的HCC肝的冷冻切片。结果通过多变量分析，肿瘤样品中的m / z 904.83和m / z 874.79分别显着较高和较低，并且被确定为甘油三酸酯（TG）16：0/18：1（9Z）/ 20：1（11Z） ）和TG 16：0/18：1（9Z）/ 18：2（9Z，12Z）使用合成化合物。TGs明显位于冷冻切片的肿瘤或非肿瘤区域。结论采用混合模式色谱柱，通过全面，可重现的G-Met方法和HDMS鉴定了HCC的新型生物标志物。UHPLC / QTOFMS和DESI-MSI的组合分析表明，TGs的不同分子种类与肿瘤分布有关，可用于表征肿瘤细胞的进程和发现前瞻性生物标志物。