Fan Ronghui, Wang Senzhen, Wu Yalan, Feng Yongli, Gao Mengke, Cao Yue, Ma Xiaoxuan, Xie Songqiang, Wang Chaojie, Gao Lei, Wang Yanming, Dai Fujun
Key Laboratory of Natural Medicine and Immuno-Engineering, Henan University, Kaifeng, Henan, China.
School of Life Sciences, Henan University, Kaifeng, Henan, China.
Phytother Res. 2023 Oct;37(10):4655-4673. doi: 10.1002/ptr.7933. Epub 2023 Aug 1.
The worldwide overall 5-year survival rate of esophageal squamous cell carcinoma (ESCC) patients is less than 20%, and novel therapeutic strategies for these patients are urgently needed. Harmine is a natural β-carboline alkaloid, which received great interest in cancer research because of its biological and anti-tumor activities. The aim of this study is to examine the effects of harmine on ESCC and its mechanism. We investigated the effects of harmine on proliferation, cell cycle, apoptosis, and tumor growth in vivo. RNA sequencing (RNA-seq), real-time PCR, and western blotting were used to detect the mechanism. Harmine inhibited ESCC cell growth in vitro and tumor growth in vivo. Differentially expressed genes in harmine-treated ESCC cells were mainly involved in protein processing in the endoplasmic reticulum (ER). Real-time PCR and western blotting confirmed harmine-induced cellular ER stress. CRISPR-Cas9 knockout of C/EBP homologous protein (CHOP) abolished harmine-induced expression of death receptor 5 and apoptosis. Harmine also induced the expression of CHOP-mediated sestrin-2, which in turn contributes to autophagosome formation via suppressing the AMP-activated protein kinase-protein kinase B-mammalian target of rapamycin signaling pathway. In conclusion, our results demonstrate that harmine inhibits the growth of ESCC through its regulation of ER stress, suggesting that it is a promising candidate for ESCC treatment.
食管鳞状细胞癌（ESCC）患者的全球总体5年生存率低于20%，因此迫切需要针对这些患者的新型治疗策略。哈尔明碱是一种天然的β-咔啉生物碱，因其生物学和抗肿瘤活性而在癌症研究中备受关注。本研究的目的是研究哈尔明碱对ESCC的影响及其作用机制。我们研究了哈尔明碱对体内增殖、细胞周期、凋亡和肿瘤生长的影响。采用RNA测序（RNA-seq）、实时PCR和蛋白质印迹法检测其作用机制。哈尔明碱在体外抑制ESCC细胞生长，在体内抑制肿瘤生长。经哈尔明碱处理的ESCC细胞中差异表达的基因主要参与内质网（ER）中的蛋白质加工。实时PCR和蛋白质印迹法证实了哈尔明碱诱导的细胞内质网应激。通过CRISPR-Cas9敲除C/EBP同源蛋白（CHOP）可消除哈尔明碱诱导的死亡受体5表达和凋亡。哈尔明碱还诱导CHOP介导的 sestrin-2表达，进而通过抑制AMP激活蛋白激酶-蛋白激酶B-雷帕霉素哺乳动物靶标信号通路促进自噬体形成。总之，我们的结果表明，哈尔明碱通过调节内质网应激来抑制ESCC的生长，提示其有望成为ESCC治疗的候选药物。
