NOX1 抑制可减轻实验性肝细胞癌中促肿瘤环境的发展。
NOX1 inhibition attenuates the development of a pro-tumorigenic environment in experimental hepatocellular carcinoma.
机构信息
Hepatology Research Unit, Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Corneel Heymanslaan 10, Blok B, 2nd floor, 9000, Ghent, Belgium.
Gut-Liver Immunopharmacology Unit, Department of Basic and Applied Medical Sciences, Faculty of Medicine and Health Sciences, Ghent University, Corneel Heymanslaan 10, Entrance 36 - Floor 3, 9000, Ghent, Belgium.
出版信息
J Exp Clin Cancer Res. 2021 Jan 23;40(1):40. doi: 10.1186/s13046-021-01837-6.
BACKGROUND
The poor prognosis of advanced HCC and limited efficacy of current systemic treatments emphasize the need for new or combined targeted therapies. The development of HCC is a multistage process in which liver injury appears in a complex microenvironment associated with oxidative stress. NOX enzymes are the main source of ROS during hepatocarcinogenesis and NOX1 in particular has shown correlation with poor prognosis of HCC patients. This study evaluates the effect of pharmacological NOX1 inhibition on the development and progression of HCC and its effect on the tumor microenvironment.
METHODS
The in vitro cytotoxic effects of the NOX1 inhibitor GKT771 (Genkyotex) on human Huh7 and Hep3B and murine Hepa1-6 HCC cell lines, the human THP1 monocyte cell line and mouse macrophages were evaluated via MTT, LDH activity and CaspGlo® assays. In order to induce in vivo HCC, male SV129 wild-type mice received weekly IP injections of diethylnitrosamine (DEN) (35 mg/kg) for 20-25 weeks. Mice were treated with vehicle or GKT771 (30 mg/kg) via oral gavage, daily or twice daily, in preventive and therapeutic studies. The liver damage was evaluated for inflammation, angiogenesis, fibrosis and HCC development via histology, RT-qPCR, multiplex analyses and ROS levels.
RESULTS
A concentration-dependent reduction in cellular activity of the human HCC cell lines without cytotoxicity was observed. GKT771 treatment reduced LPS-induced pro-inflammatory bone-marrow derived macrophage polarization. DEN injections resulted in 100 % tumor formation and the induction of HCC markers which could be reduced by twice daily dosing of GKT771 at early onset of advanced HCC. DEN-induced HCC resulted in an upregulation of pro-inflammatory, angiogenic and fibrotic markers which was less pronounced in GKT771 treated mice in all treatment regimens. In line, liver fibrosis was induced in HCC mice and this to a lesser extend upon GKT771 treatment.
CONCLUSIONS
NOX1 inhibition showed to be safe and well tolerated and was able to attenuate the induction of a pro-inflammatory, angiogenic and pro-fibrotic microenvironment suggesting that this might be a promising adjuvant therapeutic strategy in the treatment of advanced HCC.
背景
晚期 HCC 的预后较差，且当前全身治疗的疗效有限，这强调了需要新的或联合靶向治疗。肝癌的发生是一个多阶段的过程，肝损伤发生在与氧化应激相关的复杂微环境中。NOX 酶是肝癌发生过程中 ROS 的主要来源，特别是 NOX1 与 HCC 患者的预后不良相关。本研究评估了药理学 NOX1 抑制对 HCC 的发生和进展的影响及其对肿瘤微环境的影响。
方法
通过 MTT、LDH 活性和 CaspGlo®测定法评估 NOX1 抑制剂 GKT771（Genkyotex）对人 Huh7 和 Hep3B 以及鼠 Hepa1-6 HCC 细胞系、人 THP1 单核细胞系和鼠巨噬细胞的体外细胞毒性作用。为了诱导体内 HCC，雄性 SV129 野生型小鼠每周接受腹腔注射二乙基亚硝胺（DEN）（35mg/kg）20-25 周。在预防性和治疗性研究中，通过口服灌胃，每日或每日两次给予载体或 GKT771（30mg/kg）治疗。通过组织学、RT-qPCR、多重分析和 ROS 水平评估肝损伤的炎症、血管生成、纤维化和 HCC 发展。
结果
观察到人类 HCC 细胞系的细胞活性呈浓度依赖性降低，而无细胞毒性。GKT771 处理可减少 LPS 诱导的促炎骨髓来源的巨噬细胞极化。DEN 注射导致 100%肿瘤形成，并诱导 HCC 标志物，在早期晚期 HCC 时每日两次给予 GKT771 可降低这些标志物。DEN 诱导的 HCC 导致促炎、血管生成和纤维化标志物上调，在所有治疗方案中，用 GKT771 治疗的小鼠的这些标志物上调程度较低。同样，HCC 小鼠发生肝纤维化，而 GKT771 治疗则减轻了这种纤维化。
结论
NOX1 抑制显示安全且耐受良好，并能够减弱促炎、血管生成和促纤维化微环境的诱导，表明这可能是治疗晚期 HCC 的一种有前途的辅助治疗策略。
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