Shakya Rajina, Park Gyu Hwan, Joo Sang Hoon, Shim Jung-Hyun, Choi Joon-Seok
College of Pharmacy, Daegu Catholic University, Gyeongsan 38430.
College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu 41566.
Biomol Ther (Seoul). 2022 Nov 1;30(6):585-592. doi: 10.4062/biomolther.2022.121.
Treatment of triple-negative breast cancer (TNBC) has been limited due to the lack of molecular targets. In this study, we evaluated the cytotoxicity of hydroxyzine, a histamine H1 receptor antagonist in human triple-negative breast cancer BT-20 and HCC-70 cells. Hydroxyzine inhibited the growth of cells in dose- and time-dependent manners. The annexin V/propidium iodide double staining assay showed that hydroxyzine induced apoptosis. The hydroxyzine-induced apoptosis was accompanied down-regulation of cyclins and CDKs, as well as the generation of reactive oxygen species (ROS) without cell cycle arrest. The effect of hydroxyzine on the induction of ROS and apoptosis on TNBC cells was prevented by pre-treatment with ROS scavengers, N-acetyl cysteine or Mito-TEMPO, a mitochondria-targeted antioxidant, indicating that an increase in the generation of ROS mediated the apoptosis induced by hydroxyzine. Western blot analysis showed that hydroxyzine-induced apoptosis was through down-regulation of the phosphorylation of JAK2 and STAT3 by hydroxyzine treatment. In addition, hydroxyzine induced the phosphorylation of JNK and p38 MAPK. Our results indicate that hydroxyzine induced apoptosis via mitochondrial superoxide generation and the suppression of JAK2/STAT3 signaling.
由于缺乏分子靶点，三阴性乳腺癌（TNBC）的治疗一直受到限制。在本研究中，我们评估了组胺H1受体拮抗剂羟嗪对人三阴性乳腺癌BT-20和HCC-70细胞的细胞毒性。羟嗪以剂量和时间依赖性方式抑制细胞生长。膜联蛋白V/碘化丙啶双染色分析表明羟嗪诱导细胞凋亡。羟嗪诱导的细胞凋亡伴随着细胞周期蛋白和细胞周期蛋白依赖性激酶的下调，以及活性氧（ROS）的产生，且无细胞周期停滞。用ROS清除剂N-乙酰半胱氨酸或线粒体靶向抗氧化剂Mito-TEMPO预处理可阻止羟嗪对TNBC细胞ROS诱导和凋亡的影响，表明ROS生成增加介导了羟嗪诱导的细胞凋亡。蛋白质免疫印迹分析表明，羟嗪诱导的细胞凋亡是通过羟嗪处理下调JAK2和STAT3的磷酸化实现的。此外，羟嗪诱导JNK和p38丝裂原活化蛋白激酶的磷酸化。我们的结果表明，羟嗪通过线粒体超氧化物生成和JAK2/STAT3信号通路的抑制诱导细胞凋亡。
