Tsai Chun-Hao, Hsieh Pei-Wen, Lee Yi-Chen, Wang Chie-Hong, Chiu Wen-Chin, Lu Chun-Wun, Wang Yen-Yun, Hu Stephen Chu-Sung, Cheng Tain-Lu, Yuan Shyng-Shiou F
Translational Research Center, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Graduate Institute of Natural Products, School of Traditional Chinese Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan; Department of Anesthesiology, Chang Gung Memorial Hospital, Taoyuan, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Life Sci. 2017 Mar 1;172:19-26. doi: 10.1016/j.lfs.2016.12.009. Epub 2016 Dec 20.
Members of the β-nitrostyrene family are known to suppress tumor growth, with the underlying mechanisms of β-nitrostyrene remain mostly unclear. Herein, we synthesized a β-nitrostyrene derivative, 3'-hydroxy-4'-methoxy-β-methyl-β-nitrostyrene (CYT-Rx20), and explored its anticancer activities in human lung cancer cells in vitro and in vivo.
Cell viability was measured by XTT assay. Apoptosis was detected by Annexin V/PI staining. Caspase activation was determined by western blotting. ROS (reactive oxygen species), MMP (mitochondrial membrane potential) and mitochondrial mass were determined by flow cytometry. GSH level was detected by ELISA assay.
In this study, we found that CYT-Rx20 significantly reduced cell viability, accompanied by G2/M arrest in lung cancer cells. Increased protein levels of cleaved-caspase families indicated apoptotic cell death upon CYT-Rx20 treatment. Furthermore, increased level of intracellular reactive oxygen species (ROS), loss of mitochondrial membrane potential (ΔΨm), glutathione (GSH) depletion and inhibition of GSH reductase were observed after CYT-Rx20 treatment. The effects of CYT-Rx20 on cell viability and the loss of ΔΨm were significantly reversed when cells were pretreated with thiol antioxidants NAC, GSH, or 2-ME. Finally, xenograft animal study demonstrated that CYT-Rx20 significantly suppressed lung tumor growth in vivo.
Our data demonstrated that CYT-Rx20 triggered apoptotic cell death in lung cancer cells and suppressed lung tumor growth through GSH depletion, suggesting that CYT-Rx20 may have the potential to be further developed as an anticancer compound for treating lung cancer.
已知β-硝基苯乙烯家族成员可抑制肿瘤生长，但其潜在机制大多仍不清楚。在此，我们合成了一种β-硝基苯乙烯衍生物，3'-羟基-4'-甲氧基-β-甲基-β-硝基苯乙烯（CYT-Rx20），并在体外和体内研究了其对人肺癌细胞的抗癌活性。
通过XTT法检测细胞活力。采用Annexin V/PI染色检测细胞凋亡。通过蛋白质印迹法测定半胱天冬酶激活情况。采用流式细胞术测定活性氧（ROS）、线粒体膜电位（MMP）和线粒体质量。通过ELISA法检测谷胱甘肽（GSH）水平。
在本研究中，我们发现CYT-Rx20显著降低细胞活力，并伴有肺癌细胞的G2/M期阻滞。裂解的半胱天冬酶家族蛋白水平升高表明CYT-Rx20处理后细胞发生凋亡性死亡。此外，CYT-Rx20处理后观察到细胞内活性氧水平升高、线粒体膜电位（ΔΨm）丧失、谷胱甘肽（GSH）消耗以及谷胱甘肽还原酶受到抑制。当用硫醇抗氧化剂NAC、GSH或2-ME预处理细胞时，CYT-Rx20对细胞活力和ΔΨm丧失的影响显著逆转。最后，异种移植动物研究表明CYT-Rx20在体内显著抑制肺癌肿瘤生长。
我们的数据表明CYT-Rx20通过消耗GSH触发肺癌细胞的凋亡性死亡并抑制肺癌肿瘤生长，这表明CYT-Rx20可能有潜力进一步开发成为治疗肺癌的抗癌化合物。
