Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung.
Graduate Institute of Natural Products, School of Traditional Chinese Medicine, and Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University.
Int J Gynecol Cancer. 2017 Sep;27(7):1306-1317. doi: 10.1097/IGC.0000000000001033.
The β-nitrostyrene family has been reported to possess anticancer properties. However, the anticancer activity of β-nitrostyrenes on cervical cancer cells and the underlying mechanisms involved remain unexplored. In this study, a β-nitrostyrene derivative CYT-Rx20 (3'-hydroxy-4'-methoxy-β-methyl-β-nitrostyrene) was synthesized, and its anticancer activity on cervical cancer cells and the mechanisms involved were investigated.
The effect of CYT-Rx20 on human cervical cancer cell growth was evaluated using cell viability assay. Reactive oxygen species (ROS) generation and annexin V staining were detected by flow cytometry. The protein expression levels of cleaved caspase-3, cleaved caspase-9, cleaved poly (ADPribose) polymerase, γH2AX, β-catenin, Vimentin, and Twist were measured by Western blotting. DNA double-strand breaks were determined by γ-H2AX foci formation and neutral comet assay. Migration assay was used to determine cancer cell migration. Nude mice xenograft was used to investigate the antitumor effects of CYT-Rx20 in vivo.
CYT-Rx20 induced cytotoxicity in cervical cancer cells by promoting cell apoptosis via ROS generation and DNA damage. CYT-Rx20-induced cell apoptosis, ROS generation, and DNA damage were reversed by thiol antioxidants. In addition, CYT-Rx20 inhibited cervical cancer cell migration by regulating the expression of epithelial-to-mesenchymal transition markers. In nude mice, CYT-Rx20 inhibited cervical tumor growth accompanied by increased expression of DNA damage marker γH2AX and decreased expression of mesenchymal markers β-catenin and Twist.
CYT-Rx20 inhibits cervical cancer cells in vitro and in vivo and has the potential to be further developed into an anti-cervical cancer drug clinically.
已有报道称β-亚硝基苯乙烯家族具有抗癌特性。然而，β-亚硝基苯乙烯类化合物对宫颈癌的抗癌活性及其潜在的作用机制尚未得到探索。本研究合成了一种β-亚硝基苯乙烯衍生物 CYT-Rx20（3'-羟基-4'-甲氧基-β-甲基-β-亚硝基苯乙烯），并研究了其对宫颈癌的抗癌活性及其作用机制。
采用细胞活力测定法评估 CYT-Rx20 对人宫颈癌的生长抑制作用。采用流式细胞术检测活性氧（ROS）的产生和 Annexin V 染色。采用 Western blot 检测 cleaved caspase-3、cleaved caspase-9、cleaved poly（ADP-ribose）polymerase、γH2AX、β-catenin、Vimentin 和 Twist 的蛋白表达水平。通过γ-H2AX 焦点形成和中性彗星试验测定 DNA 双链断裂。采用迁移实验检测癌细胞迁移。裸鼠异种移植实验用于研究 CYT-Rx20 在体内的抗肿瘤作用。
CYT-Rx20 通过 ROS 生成和 DNA 损伤促进细胞凋亡，诱导宫颈癌细胞的细胞毒性。硫醇抗氧化剂可逆转 CYT-Rx20 诱导的细胞凋亡、ROS 生成和 DNA 损伤。此外，CYT-Rx20 通过调节上皮间质转化标志物的表达抑制宫颈癌的迁移。在裸鼠中，CYT-Rx20 抑制了宫颈肿瘤的生长，同时伴随着 DNA 损伤标志物 γH2AX 的表达增加和间充质标志物β-catenin 和 Twist 的表达减少。
CYT-Rx20 在体外和体内抑制宫颈癌，并有可能进一步开发为临床上用于治疗宫颈癌的药物。
