慢性丙型肝炎病毒感染的肝癌细胞中外在凋亡途径的改变调节增强了对马帕他木单抗诱导凋亡的敏感性。
Altered regulation of extrinsic apoptosis pathway in HCV-infected HCC cells enhances susceptibility to mapatumumab-induced apoptosis.
机构信息
LIR, NIAID, NIH, DHHS, Bethesda, USA.
出版信息
Hepatol Res. 2009 Dec;39(12):1178-89. doi: 10.1111/j.1872-034X.2009.00568.x. Epub 2009 Sep 25.
BACKGROUND
Hepatitis C virus (HCV)-infected patients, including those co-infected with human immunodeficiency virus (HIV), are at increased risk of developing hepatocellular carcinoma (HCC). We evaluated the ability of agonistic human monoclonal antibodies to tumor necrosis factor-related apoptosis inducing ligand (TRAIL) receptors, mapatumumab and lexatumumab, respectively, to induce TRAIL-receptor mediated apoptosis (TRMA) in HCC (HCV-infected and -uninfected) cells and in peripheral blood cells (HIV-infected and -uninfected).
METHODS
Susceptibility to antibody-mediated TRMA was measured by caspase 3/7 activity and by confocal microscopy. Surface expression of receptors on HCV-uninfected and -infected Huh7.5 cells was measured by flow cytometry and confocal microscopy. Inhibitor of Apoptosis Protein (IAP) RNA levels were quantified by RT-PCR. DNA Microarray was performed using RNA isolated from Huh7.5 cells (HCV-infected and uninfected) using Affymetrix U133A chips.
RESULTS
Mapatumumab preferentially induces TRMA of HCV-infected Huh7.5 cells by binding to TRAIL-R1. Higher basal expression of TRAIL-R2 compared to that of TRAIL-R1 on HCV-uninfected Huh7.5 cells were observed. Lexatumumab induces TRMA of both HCV-infected and -uninfected cells by binding to TRAIL-R2. IFN-alpha has minimal effect on mapatumumab- and lexatumumab-induced TRMA. HCV infection of Huh7.5 cells up-regulates TRAIL-R1 expression and X-linked Inhibitor of apoptosis protein and survivin gene expression. Neither antibody had a pro-apoptotic effect on PBMCs from patients with HIV infection ex vivo.
CONCLUSION
Both mapatumumab and lexatumumab are excellent candidates for therapy of HCC. HCV infection of Huh7.5 cells selectively up-regulates TRAIL-R1 receptor, associated with increased susceptibility to mapatumumab-mediated TRMA. HCV infection up-regulated IAP genes, offering promise for future combination therapy using TRAIL agonists and IAP inhibitors.
背景
丙型肝炎病毒（HCV）感染患者，包括合并人类免疫缺陷病毒（HIV）感染的患者，发生肝细胞癌（HCC）的风险增加。我们评估了激动型人源单克隆抗体分别针对肿瘤坏死因子相关凋亡诱导配体（TRAIL）受体 1（mapatumumab）和 2（lexatumumab）在诱导 HCC（HCV 感染和未感染）细胞和外周血单个核细胞（HIV 感染和未感染）中 TRAIL 受体介导的凋亡（TRMA）的能力。
方法
通过 caspase 3/7 活性和共聚焦显微镜来测量对抗体介导的 TRMA 的敏感性。通过流式细胞术和共聚焦显微镜来测量 HCV 未感染和感染 Huh7.5 细胞上受体的表面表达。通过 RT-PCR 来定量凋亡抑制蛋白（IAP）RNA 水平。使用来自 Huh7.5 细胞（HCV 感染和未感染）的 RNA 进行 Affymetrix U133A 芯片 DNA 微阵列。
结果
mapatumumab 通过与 TRAIL-R1 结合，优先诱导 HCV 感染的 Huh7.5 细胞的 TRMA。在 HCV 未感染的 Huh7.5 细胞中，观察到 TRAIL-R2 的基础表达高于 TRAIL-R1。Lexatumumab 通过与 TRAIL-R2 结合，诱导 HCV 感染和未感染细胞的 TRMA。IFN-α对 mapatumumab 和 lexatumumab 诱导的 TRMA 的影响最小。HCV 感染 Huh7.5 细胞上调 TRAIL-R1 表达和 X 连锁凋亡抑制蛋白和生存素基因表达。两种抗体在体外均未对 HIV 感染患者的 PBMC 产生促凋亡作用。
结论
mapatumumab 和 lexatumumab 均是 HCC 治疗的优秀候选药物。HCV 感染 Huh7.5 细胞选择性地上调 TRAIL-R1 受体，与 mapatumumab 介导的 TRMA 敏感性增加相关。HCV 感染上调 IAP 基因，为未来使用 TRAIL 激动剂和 IAP 抑制剂联合治疗提供了希望。
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