Hu Xue, Yang Zihui, Liu Wenjing, Pan Zhaohai, Zhang Xin, Li Minjing, Liu Xiaona, Zheng Qiusheng, Li Defang
Yantai Key Laboratory of Pharmacology of Traditional Chinese Medicine in Tumor Metabolism, School of Integrated Traditional Chinese and Western Medicine, Binzhou Medical University, Yantai, China.
Key Laboratory of Xinjiang Endemic Phytomedicine Resources of Ministry of Education, School of Pharmacy, Shihezi University, Shihezi, China.
Front Oncol. 2020 Oct 16;10:558414. doi: 10.3389/fonc.2020.558414. eCollection 2020.
Existing research shows that coumaric acid (CA) can inhibit the proliferation of a variety of tumor cells . However, there are no reports on the anti-tumor effects of CA on melanoma cells. In this study, the inhibitory effects of CA on mouse melanoma B16 and human melanoma A375 cells are reported, and the related mechanisms are investigated. CCK-8 assay was used to detect the effects of CA on cell vitality, colony formation assay was used to observe the effects on cell proliferation, Hoechst 33,258 staining was used to observe the morphology of apoptotic cells, flow cytometry was used to detect the effects on apoptosis and the cell cycle, and western blot was used to measure the levels of cell cycle- and apoptosis-related signaling pathway proteins. CA significantly inhibits cell proliferation of A375 and B16 cells in a dose-dependent manner and obviously induced cell morphological changes. CA arrested A375 cells in the S phase by downregulating the cell cycle-related proteins Cyclin A and CDK2, and arrested B16 cells in the G0-G1 phase through downregulating the cell cycle-related proteins Cyclin E and CDK2. In addition, CA significantly promoted apoptosis of A375 and B16 cells. Furthermore, CA significantly upregulated the levels of Apaf1 and Bax and downregulated the levels of Bcl-2, and subsequently increased the levels of cytoplasmic cytochrome c (Cyto-c), cleaved caspase-3, and cleaved caspase-9, leading to apoptosis in A375 and B16 cells. CA can significantly inhibit the proliferation of human and mouse melanoma cells . Our research is a step in the development of anti-melanoma drugs.
现有研究表明,香豆酸(CA)可抑制多种肿瘤细胞的增殖。然而,关于CA对黑色素瘤细胞的抗肿瘤作用尚无报道。在本研究中,报道了CA对小鼠黑色素瘤B16细胞和人黑色素瘤A375细胞的抑制作用,并对相关机制进行了研究。采用CCK-8法检测CA对细胞活力的影响,采用集落形成法观察对细胞增殖的影响,采用Hoechst 33258染色观察凋亡细胞的形态,采用流式细胞术检测对细胞凋亡和细胞周期的影响,采用蛋白质免疫印迹法检测细胞周期和凋亡相关信号通路蛋白的水平。CA以剂量依赖性方式显著抑制A375和B16细胞的增殖,并明显诱导细胞形态变化。CA通过下调细胞周期相关蛋白Cyclin A和CDK2使A375细胞停滞于S期,通过下调细胞周期相关蛋白Cyclin E和CDK2使B16细胞停滞于G0-G1期。此外,CA显著促进A375和B16细胞凋亡。此外,CA显著上调Apaf1和Bax的水平,下调Bcl-2的水平,随后增加细胞质细胞色素c(Cyto-c)、裂解的caspase-3和裂解的caspase-9的水平,导致A375和B16细胞凋亡。CA可显著抑制人源和小鼠源黑色素瘤细胞的增殖。我们的研究是抗黑色素瘤药物开发的一个进展。