Kim Min Jeong, Ku Jin Mo, Hong Se Hyang, Kim Hyo In, Kwon Yun Young, Park Joon-Sang, Jung Deok Hyun, Shin Yong Cheol, Ko Seong-Gyu
Department of Science in Korean Medicine, Graduate School, Kyung Hee University, Seoul, South Korea.
Department of Preventive Medicine, College of Korean Medicine, Kyung Hee University, Seoul, South Korea.
Front Pharmacol. 2021 May 13;12:683575. doi: 10.3389/fphar.2021.683575. eCollection 2021.
Prostate cancer is the second most commonly diagnosed cancer, and prostate cancer is the second most common cause of cancer death in United States men after lung cancer. Many therapies are used to treat prostate cancer, and chemotherapy is one of the most relevant treatments. However, chemotherapy has many side effects, and repeated administration of chemotherapeutic agents leads to acquired resistance. Thus, new drugs with few side effects are needed. We investigated the molecular mechanism of action of JI017 in human prostate cancer cells. We identified an endoplasmic reticulum (ER) stress pathway that depended on the reactive oxygen species (ROS) pathway and played a crucial role in JI017-induced apoptosis. We measured cell viability by the MTS assay to determine the effect of JI017. Analysis of apoptosis, mitochondrial dysfunction, and cell cycle features was performed by flow cytometry. We used western blot and RT-PCR to measure the levels of the proteins of the unfolded protein response (UPR) pathway and apoptosis markers. Immunoprecipitation assay and transfection were used to determine the expression levels of proteins interacting with the pathways influenced by JI017 in prostate cancer cells. The anticancer effects induced by JI017 were evaluated. JI017 induced cell death that regulated apoptotic molecules and caused cell cycle arrest that inhibited the proliferation of cancer cells. Moreover, JI017 generated ROS. Accumulation of ROS caused ER stress through the PERK-eIF2α-CHOP and IRE1α-CHOP pathways. Furthermore, persistent activation of the UPR pathway induced by JI017 treatment triggered mitochondrial dysfunction, including dissipation of mitochondrial membrane potential, which activated intrinsic apoptotic pathway in human prostate cancer cells. The data indicated that N-acetyl-L-cysteine diminished apoptosis. We demonstrated that JI017 induced ER stress and cell death. Anticancer properties of JI017 in prostate cancer cells and in a human prostate cancer model involved ROS-mediated ER stress. Thus, JI017 treatment provides a new strategy for chemotherapy of prostate cancer.
前列腺癌是第二大最常被诊断出的癌症，在美国男性中，前列腺癌是仅次于肺癌的第二大常见癌症死亡原因。许多疗法被用于治疗前列腺癌，化疗是最常用的治疗方法之一。然而，化疗有许多副作用，并且重复给药化疗药物会导致获得性耐药。因此，需要副作用少的新药。我们研究了JI017在人前列腺癌细胞中的分子作用机制。我们确定了一条依赖于活性氧（ROS）途径的内质网（ER）应激途径，该途径在JI017诱导的细胞凋亡中起关键作用。我们通过MTS试验测量细胞活力以确定JI017的作用效果。通过流式细胞术分析细胞凋亡、线粒体功能障碍和细胞周期特征。我们使用蛋白质印迹法和逆转录-聚合酶链反应（RT-PCR）来测量未折叠蛋白反应（UPR）途径的蛋白质水平和凋亡标志物。免疫沉淀试验和转染用于确定与前列腺癌细胞中受JI017影响的途径相互作用的蛋白质的表达水平。评估了JI017诱导的抗癌作用。JI017诱导细胞死亡，调节凋亡分子，并导致细胞周期停滞，从而抑制癌细胞的增殖。此外，JI017产生活性氧。活性氧的积累通过PERK-eIF2α-CHOP和IRE1α-CHOP途径导致内质网应激。此外，JI017处理诱导的UPR途径的持续激活引发线粒体功能障碍，包括线粒体膜电位的消散，从而激活人前列腺癌细胞中的内源性凋亡途径。数据表明N-乙酰-L-半胱氨酸减少了细胞凋亡。我们证明了JI017诱导内质网应激和细胞死亡。JI017在前列腺癌细胞和人前列腺癌模型中的抗癌特性涉及活性氧介导的内质网应激。因此，JI017治疗为前列腺癌化疗提供了一种新策略。
