血管紧张素 II 和肿瘤坏死因子-α 通过下调 GRK2 表达来刺激 BEL-7402 细胞的生长、迁移和侵袭。

Angiotensin II and tumor necrosis factor-α stimulate the growth, migration and invasion of BEL-7402 cells via down-regulation of GRK2 expression.

机构信息

Institute of Clinical Pharmacology, Anhui Medical University, Key Laboratory of Anti-inflammatory and Immune Medicine, Ministry of Education, Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine, Hefei, Anhui, PR China; Department of Emergency Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, PR China.

出版信息

Dig Liver Dis. 2019 Feb;51(2):263-274. doi: 10.1016/j.dld.2018.06.007. Epub 2018 Jun 20.

DOI:10.1016/j.dld.2018.06.007

PMID:30712649

Abstract

PURPOSE

To investigate the effects of angiotensin II (Ang II) and tumor necrosis factor-α (TNF-α) on the biological characteristics of hepatocellular carcinoma (HCC) cells and the associated changes in G protein-coupled receptor kinase 2 (GRK2) expression.

METHODS

The mean serum levels of Ang II and TNF-α in normal subjects and patients with benign liver tumors (BLTs) and HCC were evaluated by enzyme-linked immunosorbent assay (ELISA), and liver samples from the patients with HCC and HCC mice were used to assess the protein levels of both cytokines, their major receptors and GRK2. In addition, the dynamics of Bel-7402 cells were determined with cell counting kit-8 (CCK-8) and Transwell experiments, while the levels of the primary cytokine receptors Ang II type-1 receptor (AT1R) and type-2 receptor (AT2R) as well as TNF receptor 1 (TNFR1) were detected by flow cytometry (FCM). The effects of Ang II and TNF-α on the GRK2 levels in Bel-7402 cells and on the dynamics of GRK2-knockdown HCC cells were also investigated.

RESULTS

Both cytokines independently enhanced Bel-7402 cell growth, migration and invasion by decreasing the GRK2 level. In contrast, down-regulating the GRK2 level in Bel-7402 cells suppressed these effects. No synergistic effects were discovered when Ang II and TNF-α were administered together. Furthermore, increased AT1R and TNFR1 levels stimulated HCC initiation and progression, whereas AT2R overexpression produced the opposite effect.

CONCLUSIONS

The present results suggested that Ang II and TNF-α promote Bel-7402 cell growth, migration and invasion by down-regulating GRK2 expression, and that the associated receptors AT1R, AT2R and TNFR1 participate in HCC initiation and progression.

摘要

目的

探讨血管紧张素Ⅱ（AngⅡ）和肿瘤坏死因子-α（TNF-α）对肝癌（HCC）细胞生物学特性的影响及G 蛋白偶联受体激酶 2（GRK2）表达的变化。

方法

采用酶联免疫吸附试验（ELISA）检测正常人群和良性肝肿瘤（BLT）及 HCC 患者的血清 AngⅡ和 TNF-α平均水平，并采用免疫印迹法（Western blot）检测 HCC 患者及 HCC 小鼠肝组织中两种细胞因子及其主要受体和 GRK2 的蛋白水平。此外，通过细胞计数试剂盒-8（CCK-8）和 Transwell 实验检测 Bel-7402 细胞的动态变化，采用流式细胞术（FCM）检测主要细胞因子受体血管紧张素Ⅱ 1 型受体（AT1R）和 2 型受体（AT2R）以及肿瘤坏死因子受体 1（TNFR1）的水平。还研究了 AngⅡ和 TNF-α对 Bel-7402 细胞中 GRK2 水平的影响以及对 GRK2 敲低 HCC 细胞动态变化的影响。

结果

两种细胞因子均可通过降低 GRK2 水平独立增强 Bel-7402 细胞的生长、迁移和侵袭。相反，下调 Bel-7402 细胞中的 GRK2 水平则抑制了这些作用。当 AngⅡ和 TNF-α同时给药时，未发现协同作用。此外，AT1R 和 TNFR1 水平的升高刺激 HCC 的发生和进展，而 AT2R 的过表达则产生相反的效果。