Wang Xiaoqing, Liu Yang, Wu Rongde, Guo Feng, Zhang Lijuan, Cui Mingyu, Wu Xiangyu, Zhang Yongfei, Liu Wei
Department of Pediatric Surgery, Shandong Provincial Hospital affiliated to Shandong University, Jinan, People's Republic of China.
Department of Dermatology, Shandong Provincial Qianfoshan Hospital affiliated to Shandong University, Jinan, People's Republic of China.
Onco Targets Ther. 2018 Feb 22;11:943-953. doi: 10.2147/OTT.S157480. eCollection 2018.
Malignant melanoma (MM) is a malignant tumor produced by changes in melanocytes in the skin or other organs. In the classification of skin tumor mortality, skin melanoma ranks the highest. Ubenimex, an Aminopeptidase N (APN) inhibitor, is now widely used for cancer as an adjunct therapy, conferring antitumor effects. Apoptosis and the induction of autophagy have both been found to be closely associated with tumor cell death.
In this study, the A375 and A2058 cell lines were treated with ubenimex. Cell viability was measured using the Cell Counting Kit 8 assay. Apoptosis and autophagic cell death were assessed using flow cytometry and acridine orange/ethidium bromide staining. Protein expression was assessed by Western blot analyses and immunofluorescence. Matrigel invasion and migration assays were used to examine the metastatic ability of melanoma cells.
The results revealed that ubenimex inhibited the expression of APN in melanoma cells, which may be connected with the inhibition of metastasis. In addition, it increased melanoma cell death by inducing apoptosis and autophagic cell death. This effect was accompanied by increased levels of p-JNK. Moreover, treatment with ubenimex induced protective Akt activation, and combined use of an Akt inhibitor with ubenimex provided a better effect for inducing tumor cell death.
As an effective anti-tumor drug in vitro, ubenimex might be an excellent adjunctive therapy for the treatment of melanoma, with greater effects when combined with the use of an Akt inhibitor.
恶性黑色素瘤（MM）是由皮肤或其他器官中的黑素细胞发生变化而产生的恶性肿瘤。在皮肤肿瘤死亡率分类中，皮肤黑色素瘤位居首位。乌苯美司是一种氨肽酶N（APN）抑制剂，目前作为辅助治疗广泛应用于癌症治疗，具有抗肿瘤作用。细胞凋亡和自噬的诱导均已被发现与肿瘤细胞死亡密切相关。
在本研究中，用乌苯美司处理A375和A2058细胞系。使用细胞计数试剂盒8检测法测量细胞活力。采用流式细胞术和吖啶橙/溴化乙锭染色评估细胞凋亡和自噬性细胞死亡。通过蛋白质印迹分析和免疫荧光评估蛋白质表达。使用基质胶侵袭和迁移试验检测黑色素瘤细胞的转移能力。
结果显示，乌苯美司抑制黑色素瘤细胞中APN的表达，这可能与转移抑制有关。此外，它通过诱导细胞凋亡和自噬性细胞死亡增加黑色素瘤细胞死亡。这种作用伴随着p-JNK水平的升高。此外，用乌苯美司处理可诱导保护性Akt激活，并且将Akt抑制剂与乌苯美司联合使用对诱导肿瘤细胞死亡具有更好的效果。
作为一种有效的体外抗肿瘤药物，乌苯美司可能是治疗黑色素瘤的一种优秀辅助疗法，与Akt抑制剂联合使用时效果更佳。
