Li Xiaohan, Li Bo, Li Boan, Guo Tongsheng, Sun Zhiqiang, Li Xiaoxi, Chen Lin, Chen Weijiao, Chen Peng, Mao Yuanli, Zeng Yanjun
Graduate Student Team, Medical University of PLA, Beijing, China.
China Center for Clinical Laboratory, 302 Hospital of PLA, Beijing, China.
Pathol Oncol Res. 2018 Jul;24(3):663-670. doi: 10.1007/s12253-017-0285-4. Epub 2017 Aug 21.
Hepatocellular carcinoma (HCC) is a highly lethal malignant tumor evolved from cirrhosis. It is quite significant to seek accurate, easy markers for early warning and diagnosis of HCC. Through prospective cohort follow-up study and mass spectrometry, we discovered and verified a serum marker valuable for early warning and diagnosis. Follow-up observation was performed on cirrhosis patients. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was adopted to detect the serums of patients, and the serum polypeptides with a potential value in early HCC warning and diagnosis were screened. Electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF-MS/MS) was exploited to identify these screened polypeptides. Moreover, the serum marker concentration was determined by ELISA to validate the clinical value of the serum marker. Among 109 cirrhosis patients followed up for two years, 29 patients (26.6%) finally progressed into HCC. MALDI-TOF MS shows that the concentration of a 3155.66Da polypeptide was significantly different between the patients that progressed into HCC and those not. Through MS/MS identification, it is confirmed that the polypeptide is inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4). The serum ITIH4 concentrations in two groups were measured with ELISA and compared with Alpha-fetoprotein (AFP). Results show that serum ITIH4 and AFP concentrations were negatively correlated (r=-0.263, p=0.0006), and the ITIH4 concentration had a significant intergroup difference (p=0.000). Receiver operating characteristic (ROC) curve indicates that its predictive value (area under the curve, AUC) is 0.667, superior to AFP. For the patients progressing into HCC, serum samples were separately collected when they were recruited and diagnosed as cirrhosis. Measurement on these samples reveals that ITIH4 was declining during the progression of HCC (p=0.006). By virtue of mass spectrometry, we discovered and identified a biomarker valuable for early HCC warning and diagnosis. This marker overperforms the commonly used AFP, demonstrating a bright prospect.
肝细胞癌（HCC）是一种由肝硬化发展而来的高致死性恶性肿瘤。寻找准确、简便的HCC早期预警和诊断标志物具有重要意义。通过前瞻性队列随访研究和质谱分析，我们发现并验证了一种对早期预警和诊断有价值的血清标志物。对肝硬化患者进行随访观察。采用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）检测患者血清，筛选出在HCC早期预警和诊断中具有潜在价值的血清多肽。利用电喷雾电离四极杆飞行时间串联质谱（ESI-Q-TOF-MS/MS）对筛选出的多肽进行鉴定。此外，通过酶联免疫吸附测定（ELISA）测定血清标志物浓度，以验证血清标志物的临床价值。在109例随访两年的肝硬化患者中，29例（26.6%）最终进展为HCC。MALDI-TOF MS显示，进展为HCC的患者与未进展患者之间，一种3155.66Da多肽的浓度存在显著差异。通过MS/MS鉴定，证实该多肽为α-胰蛋白酶抑制剂重链4（ITIH4）。用ELISA测定两组患者血清ITIH4浓度，并与甲胎蛋白（AFP）进行比较。结果显示，血清ITIH4浓度与AFP浓度呈负相关（r=-0.263，p=0.0006），且ITIH4浓度组间差异显著（p=0.000）。受试者工作特征（ROC）曲线表明其预测价值（曲线下面积，AUC）为0.667，优于AFP。对于进展为HCC的患者，在其被招募并诊断为肝硬化时分别采集血清样本。对这些样本的检测显示，ITIH4在HCC进展过程中呈下降趋势（p=0.006）。借助质谱分析，我们发现并鉴定了一种对HCC早期预警和诊断有价值的生物标志物。该标志物优于常用的AFP，具有广阔前景。
