Abstract：
Hepatocellular carcinoma (HCC) represents a global health concern, particularly in Southeast Asia where hepatitis B virus (HBV) infection is common. In this study, we applied tissue-based proteomics to identify novel serological proteins for HCC and validated their performance in serum specimens.
In a discovery set, liver tissue specimens of HBV-related HCC, intrahepatic cholangiocarcinoma (iCCA) and colorectal cancer with liver metastasis (CRLM) were analyzed using mass spectrometry (LTQ-Orbitrap-XL). A subset of proteins that showed highly expressed in HCC were then confirmed by Western blotting. Additionally, clinical significance of selected candidate proteins was tested in serum samples of 80 patients with HBV-related HCC, 50 patients with HBV-related liver cirrhosis and 30 healthy controls.
Based on LTQ-Orbitrap-XL mass spectrometer, various differentially expressed proteins (DEPs) between tumor and adjacent non-tumor tissues were identified. These included 77 DEPs for HCC, 77 DEPs for iCCA and 55 DEPs for CRLM. Among selected candidate proteins, annexin A2 and cathepsin D were confirmed to be overexpressed in HCC tissue by Western blot analysis. In a validate cohort, serum cathepsin D level, but not annexin A2, was significantly higher in HCC compared with the non-HCC groups. Serum cathepsin D level was also positively correlated with tumor size and tumor stage. Additionally, the combined assay of serum cathepsin D and alpha-fetoprotein had a high sensitivity in detecting early HCC (83%) and intermediate/advanced HCC (96%). Moreover, patients with low serum cathepsin D (<305 ng/mL) displayed significantly better overall survival than those whose serum levels were high (≥305 ng/mL).
Proteomics and subsequent validation revealed cathepsin D as a novel biomarker for HCC. Apart from its diagnostic role, serum cathepsin D might also serve as a prognostic biomarker of HCC. Additional large-scale studies are needed to verify our findings.
In a discovery set, liver tissue specimens of HBV-related HCC, intrahepatic cholangiocarcinoma (iCCA) and colorectal cancer with liver metastasis (CRLM) were analyzed using mass spectrometry (LTQ-Orbitrap-XL). A subset of proteins that showed highly expressed in HCC were then confirmed by Western blotting. Additionally, clinical significance of selected candidate proteins was tested in serum samples of 80 patients with HBV-related HCC, 50 patients with HBV-related liver cirrhosis and 30 healthy controls.
Based on LTQ-Orbitrap-XL mass spectrometer, various differentially expressed proteins (DEPs) between tumor and adjacent non-tumor tissues were identified. These included 77 DEPs for HCC, 77 DEPs for iCCA and 55 DEPs for CRLM. Among selected candidate proteins, annexin A2 and cathepsin D were confirmed to be overexpressed in HCC tissue by Western blot analysis. In a validate cohort, serum cathepsin D level, but not annexin A2, was significantly higher in HCC compared with the non-HCC groups. Serum cathepsin D level was also positively correlated with tumor size and tumor stage. Additionally, the combined assay of serum cathepsin D and alpha-fetoprotein had a high sensitivity in detecting early HCC (83%) and intermediate/advanced HCC (96%). Moreover, patients with low serum cathepsin D (<305 ng/mL) displayed significantly better overall survival than those whose serum levels were high (≥305 ng/mL).
Proteomics and subsequent validation revealed cathepsin D as a novel biomarker for HCC. Apart from its diagnostic role, serum cathepsin D might also serve as a prognostic biomarker of HCC. Additional large-scale studies are needed to verify our findings.
摘要：
肝细胞癌（HCC）代表了全球健康问题，特别是在东南亚，乙肝病毒（HBV）感染是常见的。在这项研究中,我们应用基于组织的蛋白质组学来鉴定HCC的新型血清学蛋白,并验证其在血清标本中的性能.
在发现集中，HBV相关HCC的肝组织标本，使用质谱(LTQ-Orbitrap-XL)分析肝内胆管癌(iCCA)和结直肠癌肝转移(CRLM).然后通过蛋白质印迹确认在HCC中显示高度表达的蛋白质子集。此外,在80例HBV相关HCC患者的血清样本中测试了所选候选蛋白的临床意义，50例HBV相关性肝硬化患者和30例健康对照。
基于LTQ-Orbitrap-XL质谱仪，鉴定了肿瘤和邻近非肿瘤组织之间的各种差异表达蛋白（DEP）。其中包括77个HCC部门，为iCCA提供77个DEP，为CRLM提供55个DEP。在选定的候选蛋白质中，Westernblot分析证实膜联蛋白A2和组织蛋白酶D在HCC组织中过表达。在验证队列中，血清组织蛋白酶D水平,但不是膜联蛋白A2，在HCC中明显高于非HCC组。血清组织蛋白酶D水平也与肿瘤大小和肿瘤分期呈正相关。此外,血清组织蛋白酶D和甲胎蛋白联合检测在检测早期HCC(83%)和中/晚期HCC(96%)方面具有较高的灵敏度.此外，低血清组织蛋白酶D(<305ng/mL)患者的总生存期明显优于高血清水平(≥305ng/mL)患者.
蛋白质组学和随后的验证揭示了组织蛋白酶D作为HCC的新型生物标志物。除了它的诊断作用，血清组织蛋白酶D也可能作为HCC的预后生物标志物。需要更多的大规模研究来验证我们的发现。
在发现集中，HBV相关HCC的肝组织标本，使用质谱(LTQ-Orbitrap-XL)分析肝内胆管癌(iCCA)和结直肠癌肝转移(CRLM).然后通过蛋白质印迹确认在HCC中显示高度表达的蛋白质子集。此外,在80例HBV相关HCC患者的血清样本中测试了所选候选蛋白的临床意义，50例HBV相关性肝硬化患者和30例健康对照。
基于LTQ-Orbitrap-XL质谱仪，鉴定了肿瘤和邻近非肿瘤组织之间的各种差异表达蛋白（DEP）。其中包括77个HCC部门，为iCCA提供77个DEP，为CRLM提供55个DEP。在选定的候选蛋白质中，Westernblot分析证实膜联蛋白A2和组织蛋白酶D在HCC组织中过表达。在验证队列中，血清组织蛋白酶D水平,但不是膜联蛋白A2，在HCC中明显高于非HCC组。血清组织蛋白酶D水平也与肿瘤大小和肿瘤分期呈正相关。此外,血清组织蛋白酶D和甲胎蛋白联合检测在检测早期HCC(83%)和中/晚期HCC(96%)方面具有较高的灵敏度.此外，低血清组织蛋白酶D(<305ng/mL)患者的总生存期明显优于高血清水平(≥305ng/mL)患者.
蛋白质组学和随后的验证揭示了组织蛋白酶D作为HCC的新型生物标志物。除了它的诊断作用，血清组织蛋白酶D也可能作为HCC的预后生物标志物。需要更多的大规模研究来验证我们的发现。
