新型乙肝表面抗原（HBsAg）突变与肝细胞癌相关，阻碍HBsAg分泌并促进体外细胞增殖。
Novel HBsAg mutations correlate with hepatocellular carcinoma, hamper HBsAg secretion and promote cell proliferation in vitro.
作者信息
Salpini Romina, Surdo Matteo, Warner Nadia, Cortese Maria Francesca, Colledge Danny, Soppe Sally, Bellocchi Maria Concetta, Armenia Daniele, Carioti Luca, Continenza Fabio, Di Carlo Domenico, Saccomandi Patrizia, Mirabelli Carmen, Pollicita Michela, Longo Roberta, Romano Sara, Cappiello Giuseppina, Spanò Alberto, Trimoulet Pascale, Fleury Herve, Vecchiet Jacopo, Iapadre Nerio, Barlattani Angelo, Bertoli Ada, Mari Terenzio, Pasquazzi Caterina, Missale Gabriele, Sarrecchia Cesare, Orecchini Elisa, Michienzi Alessandro, Andreoni Massimo, Francioso Simona, Angelico Mario, Verheyen Jens, Ceccherini-Silberstein Francesca, Locarnini Stephen, Perno Carlo Federico, Svicher Valentina
机构信息
Department of Experimental Medicine and Surgery, University of Rome "Tor Vergata" Rome, Italy.
Research and Molecular Development, Victorian Infectious Diseases Reference Laboratory, North Melbourne, Victoria, Australia.
出版信息
Oncotarget. 2017 Feb 28;8(9):15704-15715. doi: 10.18632/oncotarget.14944.
BACKGROUND
An impaired HBsAg-secretion can increase HBV oncogenic-properties. Here, we investigate genetic-determinants in HBsAg correlated with HBV-induced hepatocellular carcinoma (HCC), and their impact on HBsAg-secretion and cell-proliferation.
METHODS
This study included 128 chronically HBV-infected patients: 23 with HCC (73.9% D; 26.1% A HBV-genotype), and 105 without cirrhosis/HCC (72.4% D, 27.6% A) as reference-group. The impact of mutations on HBsAg-secretion was assessed by measuring the ratio [secreted/intracellular HBsAg] until day 5 post-transfection. The impact of mutations on cell-cycle advancement was assessed by flow-cytometry.
RESULTS
Two HBsAg mutations significantly correlated with HCC: P203Q (17.4% [4/23] in HCC vs 1.0% [1/105] in non-HCC, P=0.004); S210R (34.8% [8/23] in HCC vs 3.8% [4/105] in non-HCC, P <0.001); P203Q+S210R (17.4% [4/23] in HCC vs 0% [0/110] in non-HCC, P=0.001). Both mutations reside in trans-membrane C-terminal domain critical for HBsAg-secretion. In in-vitro experiments, P203Q, S210R and P203Q+S210R significantly reduced the ratio [secreted/intracellular HBsAg] compared to wt at each time-point analysed (P <0.05), supporting an impaired HBsAg-secretion. Furthermore, P203Q and P203Q+S210R increased the percentage of cells in S-phase compared to wt, indicating cell-cycle progression (P203Q:26±13%; P203Q+S210R:29±14%; wt:18%±9, P <0.01. Additionally, S210R increased the percentage of cells in G2/M-phase (26±8% for wt versus 33±6% for S210R, P <0.001).
CONCLUSIONS
Specific mutations in HBsAg C-terminus significantly correlate with HBV-induced HCC. They hamper HBsAg-secretion and are associated with increased cellular proliferation, supporting their involvement in HCC-development. The identification of viral genetic markers associated with HCC is critical to identify patients at higher HCC-risk that may deserve intensive liver monitoring, and/or early anti-HBV therapy.
背景
乙肝表面抗原（HBsAg）分泌受损会增加乙肝病毒（HBV）的致癌特性。在此，我们研究与HBV诱导的肝细胞癌（HCC）相关的HBsAg中的基因决定因素，及其对HBsAg分泌和细胞增殖的影响。
方法
本研究纳入128例慢性HBV感染患者：23例患有HCC（D型占73.9%；A型HBV基因型占26.1%），105例无肝硬化/HCC作为参照组（D型占72.4%，A型占27.6%）。通过测量转染后第5天的[分泌型/细胞内HBsAg]比值来评估突变对HBsAg分泌的影响。通过流式细胞术评估突变对细胞周期进程的影响。
结果
两种HBsAg突变与HCC显著相关：P203Q（HCC患者中占17.4%[4/23]，非HCC患者中占1.0%[1/105]，P = 0.004）；S210R（HCC患者中占34.8%[8/23]，非HCC患者中占3.8%[4/105]，P<0.001）；P203Q + S210R（HCC患者中占17.4%[4/23]，非HCC患者中占0%[0/110]，P = 0.001）。两种突变均位于对HBsAg分泌至关重要的跨膜C末端结构域。在体外实验中，与野生型相比，P203Q、S210R和P203Q + S210R在每个分析时间点均显著降低了[分泌型/细胞内HBsAg]比值（P<0.05），支持HBsAg分泌受损。此外，与野生型相比，P203Q和P203Q + S210R增加了S期细胞的百分比，表明细胞周期进展（P203Q：26±13%；P203Q + S210R：29±14%；野生型：18%±9，P<0.01）。此外，S210R增加了G2/M期细胞的百分比（野生型为26±8%，S210R为33±6%，P<0.001）。
结论
HBsAg C末端的特定突变与HBV诱导的HCC显著相关。它们阻碍HBsAg分泌，并与细胞增殖增加有关，支持它们参与HCC的发生发展。鉴定与HCC相关的病毒基因标志物对于识别可能需要强化肝脏监测和/或早期抗HBV治疗的HCC高风险患者至关重要。
Zotero 插件