Ubenimex inhibits cell proliferation, migration and invasion in renal cell carcinoma: The effect is autophagy-associated
Ubenimex inhibits cell proliferation, migration and invasion in renal cell carcinoma: The effect is autophagy-associated
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DOI：
10.3892/or.2014.3693
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发表时间：
2015-03-01
期刊：
影响因子：
4.2
通讯作者：
Niu, Zhihong
中科院分区：
文献类型：
--
作者：
Liu, Shuai;Xie, Fang;Niu, Zhihong
乌苯美司是一种低分子量二肽，具有抑制氨肽酶N（APN）活性、增强免疫活性细胞功能和赋予抗肿瘤作用的能力。我们试图描述乌苯美司对肾细胞癌（RCC）的作用。786-O和OS-RC-2人肾细胞癌细胞系APN表达阳性，乌苯美司降低APN活性而不影响其表达。通过生长曲线分析和WST-8增殖测定，乌苯美司以浓度依赖性方式抑制两种细胞系的增殖。伤口愈合和Matrigel侵袭试验表明，RCC细胞的迁移和侵袭也被乌苯美司显着抑制。此外，乌苯美司增加死亡率的两个RCC细胞系所确定的LDH细胞毒性试验。这种影响伴随着LC 3B水平的增加，对Caspase 3没有明显的影响;我们观察到乌苯美司治疗后，两种RCC细胞系的自噬显着增加，通过电子显微镜。此外，雷帕霉素增强乌苯美司的细胞毒性作用，而3-甲基腺嘌呤逆转的效果，表明乌苯美司的细胞毒性发生通过自噬相关的机制。为了进一步评估乌苯美司在肾细胞癌治疗中的潜在适用性，我们使用组织微阵列对76例接受根治性肾切除术的肾细胞癌患者进行了免疫组化。结果表明，APN在大多数但不是所有的RCC组织中表达，并且与正常肾组织相比，APN在RCC中的表达降低，这表明APN在RCC发展中的潜在作用。总之，这些结果表明乌苯美司抑制RCC细胞的增殖、迁移和侵袭。乌苯美司可诱导自噬，这可能与其对RCC细胞的生长停滞和细胞死亡的影响有关。
Ubenimex is a low-molecular-weight dipeptide with the ability to inhibit aminopeptidase N (APN) activity, enhance the function of immunocompetent cells and confer antitumor effects. We sought to characterize the effects of ubenimex on renal cell carcinoma (RCC). The 786-O and OS-RC-2 human RCC cell lines were positive for APN expression and ubenimex decreased APN activity without affecting the expression. Ubenimex suppressed the proliferation of both cell lines in a concentration-dependent manner, as assessed by curve growth analysis and WST-8 proliferation assay. Wound healing and Matrigel invasion assays demonstrated that the migration and invasion of the RCC cells were also markedly suppressed by ubenimex. Furthermore, ubenimex increased the mortality of both RCC cell lines as determined by the LDH cytotoxicity assay. This affect was accompanied by increased levels of LC3B with no apparent effect on Caspase3; and we observed that autophagy increased significantly after ubenimex treatment in both RCC cell lines by electron microscopy. Moreover, rapamycin enhanced the cytotoxic effect of ubenimex, while 3-methyladenine reversed the effect, indicating that ubenimex cytotoxicity occured through an autophagy-related mechanism. To further assess the potential applicability of ubenimex in the treatment of RCC, we performed immunohistochemistry using tissue microarrays representing 76 RCC patients that underwent radical nephrectomy. The results showed that APN was expressed in most, but not all of the RCC tissues and that the expression was reduced in RCC as compared to the normal kidney tissues, suggesting a potential role for APN in RCC development. Collectively, these results indicated that ubenimex inhibits proliferation, migration and invasion of RCC cells. Ubenimex may induce autophagy, which may be associated with its effect on the growth arrest and the cell death of RCC cells.