Department of Radiotherapy and Oncology, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
Department of Radiology, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
J Gene Med. 2023 Sep;25(9):e3530. doi: 10.1002/jgm.3530. Epub 2023 May 19.
Colorectal cancer (CRC) is the third most common malignancy and a major cause of cancer-related deaths. Peptidyl arginine deiminase 4 (PAD4 or PADI4) is expressed in neutrophils that, when activated, can drive the formation of neutrophil extracellular traps (NETs). PAD4 has been found to be upregulated in CRC patients and to predict a poor prognosis. This study is aimed at exploring the role of PAD4 inhibitor (GSK484) in NET formation and radioresistance in CRC.
Reverse transcriptase quantitative polymerase chain reaction and western blotting were used to measure PAD4 expression in CRC tissues and cells. GSK484, an inhibitor of PAD4, was investigated in the following functional assays in vitro: western blotting, clonogenic survival, colony formation, TUNEL, flow cytometry and transwell assays. Nude mouse xenograft models were applied to evaluate the effect of GSK484 on tumor growth in CRC in vivo. The formation of NETs influenced by GSK484 was also investigated.
We showed upregulation of PAD4 mRNA and protein in CRC tissues and cells. High expression of PAD4 was related to a poor prognosis in CRC patients. GSK484 treatment promoted the radiosensitivity of CRC cells and induced cell death by promoting DNA double-strand breaks. Rescue experiments further verified that GSK484 inhibited the effects of PAD4 overexpression in irradiated CRC cells. Moreover, GSK484 injection strengthened the radiosensitivity of CRC and inhibited NET formation in vivo.
PAD4 inhibitor GSK484 promotes the radiosensitivity of CRC and inhibits NET formation in vivo and in vitro.
结直肠癌（CRC）是第三大常见恶性肿瘤，也是癌症相关死亡的主要原因。肽基精氨酸脱亚氨酶 4（PAD4 或 PADI4）在中性粒细胞中表达，当被激活时，可驱动中性粒细胞胞外诱捕网（NETs）的形成。已经发现 PAD4 在 CRC 患者中上调，并预测预后不良。本研究旨在探讨 PAD4 抑制剂（GSK484）在 CRC 中 NET 形成和放射抵抗中的作用。
逆转录定量聚合酶链反应和 Western blot 用于测量 CRC 组织和细胞中的 PAD4 表达。GSK484，一种 PAD4 的抑制剂，在体外的以下功能测定中进行了研究：Western blot、集落形成、克隆形成、TUNEL、流式细胞术和 Transwell 测定。裸鼠异种移植模型用于评估 GSK484 在 CRC 体内对肿瘤生长的影响。还研究了 GSK484 对 NET 形成的影响。
我们显示 PAD4 mRNA 和蛋白在 CRC 组织和细胞中上调。PAD4 的高表达与 CRC 患者的预后不良有关。GSK484 治疗通过促进 DNA 双链断裂，促进 CRC 细胞的放射敏感性并诱导细胞死亡。挽救实验进一步证实，GSK484 抑制了照射后 CRC 细胞中 PAD4 过表达的作用。此外，GSK484 注射增强了 CRC 的放射敏感性并抑制了体内 NET 的形成。
PAD4 抑制剂 GSK484 促进 CRC 的放射敏感性并抑制体内和体外的 NET 形成。
