基于甲醛固定石蜡包埋组织的非肝硬化和肝硬化乙肝相关肝细胞癌的差异蛋白质组学研究  

Differential Proteome Analysis of Hepatitis B Virus-related Hepatocellular Carcinoma with and without Cirrhosis from Formalin-fixed and Paraffin-embedded Tissue Samples

作　　者：盛想英 陆豪杰[1,2] SHENG Xiang-Ying;LU Hao-Jie(Department of Chemistry,Fudan University,Shanghai 200433,China;Institutes of Biomedical Sciences and NHC Key Laboratory of Glycoconjugates Research,Fudan University,Shanghai 200433,China)

机构地区：[1]复旦大学化学系,上海200433 [2]复旦大学生物医学研究院和卫健委糖复合物重点实验室,上海200433

出　　处：《分析化学》2021年第9期1478-1487,共10页Chinese Journal of Analytical Chemistry

基　　金：国家重点研发计划项目(No.2016YFA0501300);国家自然科学基金项目(No.21974025)资助。

摘　　要：肝细胞癌是起源于肝细胞的原发性肿瘤,其中30%由乙型肝炎病毒诱发。在肝细胞癌患者中,有和无肝硬化的比例约为4∶1,相关的肝癌发生机制的区别尚不明确。本研究以非肝硬化和肝硬化乙肝相关肝细胞癌患者的甲醛固定石蜡包埋肝脏肿瘤组织为对象,采用基于液相色谱-串联质谱联用(LC-MS/MS)的非标记定量方法分析差异蛋白质组,共鉴定出4083种蛋白质,其中,具有统计学差异的蛋白质283种。通过免疫组化实验,发现HSD17B13、SOD3和CXCL12在非肝硬化乙肝相关肝细胞癌组中的表达水平高于肝硬化乙肝相关肝细胞癌组,GPD2、CRTAP和CD276则正相反。本研究筛选出6种潜在的生物标志物,有望辅助非肝硬化和肝硬化乙肝相关肝细胞癌患者的机制探究和针对性治疗。生物信息学分析表明,相对于肝硬化乙肝相关肝细胞癌组,非肝硬化乙肝相关肝细胞癌组胆固醇稳态、脂肪形成通路被显著富集,并且潜在标志物提示PI3K/AKT/MMPs和Wnt/β-catenin通路均有可能与这两类的患者中不同的癌症发生过程相关,为非肝硬化和肝硬化乙肝相关肝细胞癌的癌症发生机制研究提供了基础数据和新思路。Hepatocellular carcinoma(HCC)is the fourth leading cause of deaths among malignancies,of which 30%is caused by hepatitis B virus(HBV).Most of HCC patients are associated with cirrhosis,while 20%occurs in non-cirrhotic liver.Between HBV-related HCC patients with cirrhosis(C-HBV-HCC)and without cirrhosis(NC-HBV-HCC),the mechanism of HCC is different and not yet clear.This study performed differential proteome analysis of NC-HBV-HCC and C-HBV-HCC from formalin-fixed and paraffin-embedded(FFPE)samples,using LC-MS/MS coupled with label-free quantitation.A total of 4083 proteins were identified,and 283 of them were differentially expressed between these two groups.Six proteins were validated by immunohistochemistry,among which HSD17B13/SOD3/CXCL12 were up-regulated in NC-HBV-HCC group,while the expression level of GPD2/CRTAP/CD276 were higher in C-HBV-HCC group.Overall,6 proteins were validated as biomarker candidates for guiding mechanism research of HBV-related HCC patients with and without cirrhosis,showing the potential to assist clinical management of these two patient types.Bioinformatic analyses showed that cholesterol homeostasis and adipogenesis pathways were upregulated in NC-HBV-HCC compared with C-HBV-HCC,and the 6 biomarker candidates suggested that PI3K/Akt/MMPs and Wnt/β-catenin pathways might be associated with different cancer progresses in NC-HBV-HCC and C-HBV-HCC,providing data and new ideas for the exploration of the tumorigenesis of HBV-related HCC with and without cirrhosis.

关 键 词：甲醛固定石蜡包埋 差异蛋白质组学 非肝硬化 肝硬化 乙型肝炎病毒 肝细胞癌 生物标志物 

分 类 号：R735.7[医药卫生—肿瘤]