Lab of Chemical Biology and Molecular Drug Design, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, China.
Lab of Molecular Immunology, Virus Inspection Department, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China.
Life Sci. 2021 Mar 15;269:119066. doi: 10.1016/j.lfs.2021.119066. Epub 2021 Jan 16.
This study aimed to investigate the effect of previously synthesized 4,5-diazafluorene derivative (14c) on γδ T cell-mediated cytotoxicity against renal cell carcinoma (RCC).
A real-time cell analyzer monitored cell proliferation, and Cell Counting Kit-8 determined cell viability. A reverse transcription-polymerase chain reaction analyzed gene expression, and protein expression was determined by cellular immunofluorescence analysis and Western blot.
The compound 14c induced the expression of immunomodulatory molecules, such as natural killer group 2, member D ligands (NKG2DLs), fibroblast-associated (Fas) death receptor, and tumor necrosis factor-related apoptosis-inducing ligand receptors (TRAILRs) in RCC. In addition, 14c induced DNA damage responses in RCC. Blocking DNA damage by KU-55933 reduced the effect of γδ T cells on 14c-treated RCC, suggesting that DNA damage responses were involved in the augmentation of γδ T cell-mediated cytotoxicity. Treating 786-O cells with a nitrogen-containing bisphosphonate prodrug further enhanced the anti-tumor effect of γδ T cell plus 14c combination treatment.
The present evidence indicates that 14c induced DNA damage responses in RCC and augmented γδ T cell-mediated cytotoxicity primarily through NKG2D/NKG2DLs pathways, suggesting potential cancer immunotherapy for harnessing γδ T cells and small compounds that induce DNA damage responses.
本研究旨在探讨先前合成的 4,5-二氮杂芴衍生物（14c）对 γδ T 细胞介导的肾细胞癌（RCC）细胞毒性的影响。
实时细胞分析仪监测细胞增殖，细胞计数试剂盒-8 检测细胞活力。逆转录-聚合酶链反应分析基因表达，细胞免疫荧光分析和 Western blot 检测蛋白表达。
该化合物 14c 诱导 RCC 表达免疫调节分子，如自然杀伤组 2，成员 D 配体（NKG2DLs）、成纤维细胞相关（Fas）死亡受体和肿瘤坏死因子相关凋亡诱导配体受体（TRAILRs）。此外，14c 在 RCC 中诱导 DNA 损伤反应。KU-55933 阻断 DNA 损伤可降低 γδ T 细胞对 14c 处理的 RCC 的作用，表明 DNA 损伤反应参与了 γδ T 细胞介导的细胞毒性增强。用氮杂双膦酸酯前药处理 786-O 细胞进一步增强了 γδ T 细胞加 14c 联合治疗的抗肿瘤作用。
目前的证据表明，14c 在 RCC 中诱导 DNA 损伤反应，并主要通过 NKG2D/NKG2DLs 途径增强 γδ T 细胞介导的细胞毒性，提示利用 γδ T 细胞和诱导 DNA 损伤反应的小分子进行癌症免疫治疗的潜力。
