Department of Pediatric Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China.
Department of Dermatology, Shandong Provincial Qianfoshan Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
Mol Med Rep. 2018 Mar;17(3):4531-4539. doi: 10.3892/mmr.2018.8402. Epub 2018 Jan 9.
Bladder cancer, a common urinary tract tumor, has high mortality and recurrence rates associated with metastasis. Aminopeptidase N (APN) expression and metastasis have been indicated to be associated with one another. Ubenimex may function as an APN inhibitor to inhibit the degradation of the extracellular matrix during tumorigenesis. Furthermore, APN has been widely used as an adjuvant therapy for the treatment of tumors; however, little information is available regarding the impact of ubenimex on patients. Autophagy is suggested to be important in the transformation and progression of cancer. Additionally, apoptosis, which leads to the rapid demolition of cellular organelles and structures, has also been suggested as an important factor. Thus, the present study investigated the role of ubenimex in inhibiting migration and invasion by downregulating APN expression levels to induce autophagic cell death and apoptosis in bladder cancer cells. RT112 and 5637 cell lines were treated with varying doses of ubenimex. Cell viability was measured by CCK8 colorimetry and flow cytometry. Using fluorescence microscopy, autophagic cell death was assessed using acridine orange/ethidium bromide staining. Furthermore, apoptotic cell death was assessed using flow cytometry and Trypan blue staining was used to evaluate the cell death rate. Protein expression was determined by western blot analysis. Matrigel invasion assays were exploited to assess the invasion capabilities of 5637 cells. Wound‑healing migration assays and Matrigel migration assays were exploited to assess the migratory abilities of 5637 cells. Treatment with ubenimex was accompanied by decreased Akt expression, indicating that ubenimex may have similar functions to Akt inhibitors. Results also indicated that ubenimex inhibited cell migration and invasion in bladder cancer cells. Furthermore, ubenimex also induced autophagic cell death and apoptosis, which suggested that mixed programmed cell death occurred in ubenimex‑treated bladder cancer cells. The results from the present study suggest that ubenimex may be a potential adjuvant therapy for the treatment of bladder cancer.
膀胱癌是一种常见的泌尿系统肿瘤，其死亡率和复发率较高，与转移有关。氨肽酶 N（APN）的表达与转移之间存在关联。乌苯美司可能作为 APN 抑制剂发挥作用，抑制肿瘤发生过程中外细胞基质的降解。此外，APN 已广泛用于肿瘤的辅助治疗；然而，关于乌苯美司对患者的影响的信息很少。自噬在癌症的转化和进展中具有重要作用。此外，细胞凋亡会导致细胞细胞器和结构的迅速破坏，也被认为是一个重要因素。因此，本研究探讨了乌苯美司通过下调 APN 表达水平抑制迁移和侵袭，诱导膀胱癌细胞自噬性细胞死亡和细胞凋亡的作用。用不同剂量的乌苯美司处理 RT112 和 5637 细胞系。通过 CCK8 比色法和流式细胞术测量细胞活力。使用荧光显微镜，通过吖啶橙/溴化乙锭染色评估自噬性细胞死亡。此外，通过流式细胞术评估细胞凋亡，使用台盼蓝染色评估细胞死亡率。通过 Western blot 分析测定蛋白质表达。利用 Matrigel 侵袭实验评估 5637 细胞的侵袭能力。利用划痕愈合迁移实验和 Matrigel 迁移实验评估 5637 细胞的迁移能力。乌苯美司处理伴随着 Akt 表达的降低，表明乌苯美司可能具有与 Akt 抑制剂相似的功能。结果还表明，乌苯美司抑制膀胱癌细胞的迁移和侵袭。此外，乌苯美司还诱导自噬性细胞死亡和细胞凋亡，提示乌苯美司处理的膀胱癌细胞发生混合程序性细胞死亡。本研究的结果表明，乌苯美司可能是膀胱癌治疗的一种潜在辅助治疗方法。
