Amplification of DDR2 mediates sorafenib resistance through NF-kappa B/c-Rel signaling in hepatocellular carcinoma
Amplification of DDR2 mediates sorafenib resistance through NF-kappa B/c-Rel signaling in hepatocellular carcinoma
复制标题
肝细胞癌中 DDR2 的扩增通过 NF-κ B/c-Rel 信号传导介导索拉非尼耐药
DOI：
10.1002/cbin.11625
复制
发表时间：
2021
影响因子：
3.9
通讯作者：
Xie Bin-Hui
中科院分区：
文献类型：
--
作者：
Liu Yu-Wen;Liu Qing-Quan;Xie Yuan-Kang;Zhang Jian-Hong;Song Cai-Xin;Wang Jian-Zhong;Xie Bin-Hui
索拉非尼是美国食品和药物管理局批准的第一种治疗晚期肝细胞癌（HCC）的全身治疗药物。然而，索拉非尼治疗经常伴随着耐药性。我们的目的是探讨索拉非尼耐药的机制，并提供可行的解决方案，以提高索拉非尼在晚期肝癌患者的反应。采用定量真实的时间PCR（qPCR）和蛋白质印迹法检测HCC组织和细胞中盘状结构域受体2（DDR2）的表达谱。使用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四氮唑溴化物、集落形成、TdT介导的dUTP缺口末端标记和流式细胞术测定来检查DDR2对索拉非尼耐药性的影响。通过荧光素酶报告基因、免疫荧光、qPCR和流式细胞术测定来评估DDR 2对核因子κ B（NF-κB）信号通路的影响。我们证明，相对于敏感组织，索拉非尼耐药HCC组织中DDR2表达显著上调。下调DDR2致敏的肝癌细胞系对索拉非尼的细胞毒性。进一步分析表明，DDR2可增加NF-κB亚基REL原癌基因的核定位，从而介导NF-κB信号传导。使用NF-κB信号传导抑制剂甲基巴多索龙阻断NF-κB信号传导，增加了HCC细胞对索拉非尼B的反应。进一步分析表明，DDR2的DNA扩增是导致肝癌DDR2过度表达的重要机制。我们的研究结果表明，DDR2是HCC患者的潜在治疗靶点，靶向DDR2代表了一种有希望的方法来增加HCC患者对索拉非尼的敏感性。
Sorafenib was the first systemic therapy approved by the Food and Drug Administration to treat advanced hepatocellular carcinoma (HCC). However, sorafenib therapy is frequently accompanied by drug resistance. We aimed to explore the mechanisms of sorafenib resistance and provide feasible solutions to increase the response to sorafenib in patients with advanced HCC. The expression profile of discoidin domain receptor 2 (DDR2) in HCC tissues and cells was detected using quantitative real‐time PCR (qPCR) and western blotting assays. The effects of DDR2 on sorafenib resistance were examined using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide, colony formation, TdT‐mediated dUTP nick end labeling, and flow cytometry assays. The effect of DDR2 on the nuclear factor kappa B (NF‐κB) signaling pathway was evaluated by luciferase reporter, immunofluorescence, qPCR and flow cytometry assays. We demonstrated that DDR2 expression was dramatically upregulated in sorafenib‐resistant HCC tissues relative to sensitive tissues. Downregulation ofDDR2sensitized HCC cell lines to sorafenib cytotoxicity. Further analysis showed that DDR2 could increase the nuclear location of REL proto‐oncogene, a NF‐κB subunit, to mediate NF‐κB signaling. Blocking NF‐κB signaling using the NF‐κB signaling inhibitor, bardoxolone methyl, increased the response of HCC cells to sorafenib. Further analysis showed that DNA amplification ofDDR2is an important mechanism leading toDDR2overexpression in HCC. Our results demonstrated that DDR2 is a potential therapeutic target in patients with HCC, and targeting DDR2 represents a promising approach to increase sorafenib sensitivity in patients with HCC.