Dipeptidyl peptidase‐8 induces sorafenib resistance via binding with c‐Rel to mediate NF‐κB signaling in hepatocellular carcinoma
Dipeptidyl peptidase‐8 induces sorafenib resistance via binding with c‐Rel to mediate NF‐κB signaling in hepatocellular carcinoma
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DOI：
10.1002/cbin.11719
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发表时间：
2021-10
影响因子：
3.9
通讯作者：
Hong-fu Liu;Yuankang Xie;Bai-Yin Zhong;Jianhong Zhang;Caiyun Song;Yuwen Liu;Yan Yang;B. Xie
中科院分区：
文献类型：
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作者：
Hong-fu Liu;Yuankang Xie;Bai-Yin Zhong;Jianhong Zhang;Caiyun Song;Yuwen Liu;Yan Yang;B. Xie
索拉非尼是治疗晚期肝细胞癌（HCC）患者的重要首选药物。成功治疗的主要障碍是索拉非尼耐药性。然而，索拉非尼耐药的机制尚不清楚。本研究旨在确定二肽基肽酶-8（DPP 8）参与索拉非尼耐药。使用定量真实的时间PCR（qPCR）和蛋白质印迹分析检测DPP 8表达。使用末端脱氧核苷酸转移酶缺口末端标记（TUNEL）、集落形成、流式细胞术、荧光素酶报告基因、免疫荧光和免疫沉淀（IP）试验检查DPP 8对索拉非尼耐药性的影响。我们发现肝癌中DPP 8 mRNA和蛋白水平显着上调。基因集富集分析（GSEA）表明DPP 8可能参与细胞凋亡调控。DPP 8的下调显著促进了HCC细胞对索拉非尼的敏感性。进一步的分析表明，DPP 8可能调节核因子κ B（NF-κB）信号传导，这一点使用荧光素酶报告基因测定得到证实。DPP 8的下调降低了NF-κB通路下游基因的表达水平。IP显示DPP 8可以与NF-κB亚基c-Rel相互作用，c-Rel是NF-κB信号传导的重要蛋白。最后，在DPP 8上调的细胞中，索拉非尼和瓦尔-boroPro的药物组合诱导的HCC细胞死亡率高于单独索拉非尼。我们的研究结果表明，使用抑制剂瓦尔‐boroPro可能是一种有前途的方法，以提高索拉非尼敏感性在晚期肝癌。
Sorafenib is the important first‐standard drug for patients with advanced hepatocellular carcinoma (HCC). A major obstacle to successful treatment is sorafenib resistance. However, the mechanism of sorafenib resistance is unclear. The present study aimed to determine the involvement of dipeptidyl peptidase‐8 (DPP8) in sorafenib resistance. DPP8 expression was detected using quantitative real‐time PCR (qPCR) and western blot analysis. The effect of DPP8 on sorafenib resistance was examined using terminal deoxynulceotidyl transferase nick‐end‐labeling (TUNEL), colony formation, flow cytometry, luciferase reporter, immunofluorescence, and immunoprecipitation (IP) assays. We found that DPP8 mRNA and protein levels were dramatically upregulated in HCC. Gene set enrichment analysis (GSEA) illustrated that DPP8 might be involved in apoptosis regulation. Downregulation of DPP8 substantially promoted the sensitivity of HCC cells to sorafenib. Further analysis showed that DPP8 might regulate nuclear factor kappa B (NF‐κB) signaling, which was confirmed using a luciferase reporter assay. Downregulation of DPP8 decreased the expression levels of downstream genes of the NF‐κB pathway. IP showed that DPP8 can interact with NF‐κB subunit c‐Rel, an important protein of NF‐κB signaling. Finally, a drug combination of sorafenib and Val‐boroPro induced higher mortality of HCC cells than sorafenib alone in DPP8‐upregulated cells. Our findings indicated that using the inhibitor Val‐boroPro might be a promising method to enhance sorafenib sensitivity in advanced HCC.