Consun Chinese Medicines Research Centre for Renal Diseases, School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China; Center for Cancer and Inflammation Research, School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China; Research and Development Centre for Natural Health Products, HKBU Shenzhen Research Institute and Continuing Education, Shenzhen, China.
Center for Cancer and Inflammation Research, School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China.
Phytomedicine. 2019 Nov;64:153065. doi: 10.1016/j.phymed.2019.153065. Epub 2019 Aug 2.
Melanoma is a lethal cancer. NF-κB has been validated as a molecular target for melanoma treatment. Current therapies for melanoma have limitations. Novel targeted therapeutics are needed. Arnicolide D (Ar-D), a sesquiterpene lactone isolated from the dried whole plant of Centipeda minima (L.) A. Br. et Aschers., has been reported to inhibit NF-κB activity in colorectal cancer cells.
To investigate the anti-melanoma effects of Ar-D in vitro and in vivo; and to determine whether Ar-D inhibits the NF-κB pathway in melanoma cells.
A B16F10 allograft mouse model and two melanoma cell lines (A375 and B16F10) were used to investigate the anti-melanoma effects of Ar-D in vivo and in vitro. Dacarbazine was used as a positive control. Cell viability was assessed by MTT and crystal violet staining assays. Cell cycle arrest and apoptosis were analyzed by flow cytometry. Protein levels were determined by Immunoblotting.
In vivo assays showed that the average tumor weight in Ar-D-treated group (4 mg/kg, i.p, 15 days) was reduced by 53.7%, when compared with the control group. In vitro studies demonstrated that Ar-D reduced cell viability, induced G2/M cell cycle arrest and apoptosis, elevated levels of cell cycle regulatory proteins p53 and p21, and lowered levels of G2/M checkpoint proteins Cdc2 and Cyclin B1 in melanoma cells. Mechanistically, Ar-D inhibited the activity of IKKα/β, the degradation of IκBα, and the phosphorylation and expression of NF-κB p65 in melanoma cells.
Ar-D has anti-melanoma effects, and inhibition of the IKK/IκBα/NF-κB p65 pathway is involved in the effects.
黑色素瘤是一种致命的癌症。NF-κB 已被验证为治疗黑色素瘤的分子靶标。目前治疗黑色素瘤的方法存在局限性。需要新的靶向治疗药物。arnicolide D（Ar-D）是从 Centipeda minima（L.）A. Br. et Aschers. 的干燥全植物中分离得到的一种倍半萜内酯，已被报道可抑制结直肠癌细胞中的 NF-κB 活性。
研究 Ar-D 在体外和体内的抗黑色素瘤作用；并确定 Ar-D 是否抑制黑色素瘤细胞中的 NF-κB 途径。
使用 B16F10 同种异体移植小鼠模型和两种黑色素瘤细胞系（A375 和 B16F10）在体内和体外研究 Ar-D 的抗黑色素瘤作用。使用达卡巴嗪作为阳性对照。通过 MTT 和结晶紫染色测定法评估细胞活力。通过流式细胞术分析细胞周期停滞和细胞凋亡。通过免疫印迹法测定蛋白质水平。
体内实验表明，与对照组相比，Ar-D 治疗组（4mg/kg，腹腔注射，15 天）的平均肿瘤重量减少了 53.7%。体外研究表明，Ar-D 降低了细胞活力，诱导 G2/M 细胞周期停滞和细胞凋亡，上调细胞周期调节蛋白 p53 和 p21 的水平，并降低黑色素瘤细胞中 G2/M 检查点蛋白 Cdc2 和 Cyclin B1 的水平。在机制上，Ar-D 抑制了 IKKα/β 的活性、IκBα 的降解以及 NF-κB p65 的磷酸化和表达。
Ar-D 具有抗黑色素瘤作用，抑制 IKK/IκBα/NF-κB p65 途径参与了其作用。
